130 results on '"Sandow, Jarrod J."'
Search Results
2. Ubiquitylation of RIPK3 beyond-the-RHIM can limit RIPK3 activity and cell death
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Frank, Daniel, Garnish, Sarah E., Sandow, Jarrod J., Weir, Ashley, Liu, Lin, Clayer, Elise, Meza, Lizeth, Rashidi, Maryam, Cobbold, Simon A., Scutts, Simon R., Doerflinger, Marcel, Anderton, Holly, Lawlor, Kate E., Lalaoui, Najoua, Kueh, Andrew J., Eng, Vik Ven, Ambrose, Rebecca L., Herold, Marco J., Samson, Andre L., Feltham, Rebecca, Murphy, James M., Ebert, Gregor, Pearson, Jaclyn S., and Vince, James E.
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- 2022
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3. SFPQ-ABL1 and BCR-ABL1 use different signaling networks to drive B-cell acute lymphoblastic leukemia
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Brown, Lauren M., Hediyeh-zadeh, Soroor, Sadras, Teresa, Huckstep, Hannah, Sandow, Jarrod J., Bartolo, Ray C., Kosasih, Hansen J., Davidson, Nadia M., Schmidt, Breon, Bjelosevic, Stefan, Johnstone, Ricky, Webb, Andrew I., Khaw, Seong L., Oshlack, Alicia, Davis, Melissa J., and Ekert, Paul G.
- Published
- 2022
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4. Mapping the IMiD‐dependent cereblon interactome using BioID‐proximity labelling.
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Costacurta, Matteo, Sandow, Jarrod J., Maher, Belinda, Susanto, Olivia, Vervoort, Stephin J., Devlin, Jennifer R., Garama, Daniel, Condina, Mark R., Steele, Joel R., Kahrood, Hossein V., Gough, Daniel, Johnstone, Ricky W., and Shortt, Jake
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UBIQUITIN ligases , *PROTEASOME inhibitors , *TRANSCRIPTION factors , *MULTIPLE myeloma , *CELL analysis , *MYOSIN , *CD38 antigen - Abstract
Immunomodulatory imide drugs (IMiDs) are central components of therapy for multiple myeloma (MM). IMiDs bind cereblon (CRBN), an adaptor for the CUL4‐DDB1‐RBX1 E3 ligase to change its substrate specificity and induce degradation of ‘neosubstrate’ transcription factors that are essential to MM cells. Mechanistic studies to date have largely focussed on mediators of therapeutic activity and insight into clinical IMiD toxicities is less developed. We adopted BioID2‐dependent proximity labelling (BioID2‐CRBN) to characterise the CRBN interactome in the presence and absence of various IMiDs and the proteasome inhibitor, bortezomib. We aimed to leverage this technology to further map CRBN interactions beyond what has been achieved by conventional proteomic techniques. In support of this approach, analysis of cells expressing BioID2‐CRBN following IMiD treatment displayed biotinylation of known CRBN interactors and neosubstrates. We observed that bortezomib alone significantly modifies the CRBN interactome. Proximity labelling also suggested that IMiDs augment the interaction between CRBN and proteins that are not degraded, thus designating ‘neointeractors’ distinct from previously disclosed ‘neosubstrates’. Here we identify Non‐Muscle Myosin Heavy Chain IIA (MYH9) as a putative CRBN neointeractor that may contribute to the haematological toxicity of IMiDs. These studies provide proof of concept for proximity labelling technologies in the mechanistic profiling of IMiDs and related E3‐ligase‐modulating drugs. [ABSTRACT FROM AUTHOR]
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- 2024
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5. Human RIPK3 C-lobe phosphorylation is essential for necroptotic signaling
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Meng, Yanxiang, Horne, Christopher R., Samson, Andre L., Dagley, Laura F., Young, Samuel N., Sandow, Jarrod J., Czabotar, Peter E., and Murphy, James M.
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- 2022
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6. Multimodal regulation of encystation in Giardia duodenalis revealed by deep proteomics
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Balan, Balu, Emery-Corbin, Samantha J., Sandow, Jarrod J., Ansell, Brendan Robert E., Tichkule, Swapnil, Webb, Andrew I., Svärd, Staffan G., and Jex, Aaron R.
- Published
- 2021
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7. Identification of MLKL membrane translocation as a checkpoint in necroptotic cell death using Monobodies
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Petrie, Emma J., Birkinshaw, Richard W., Koide, Akiko, Denbaum, Eric, Hildebrand, Joanne M., Garnish, Sarah E., Davies, Katherine A., Sandow, Jarrod J., Samson, Andre L., Gavin, Xavier, Fitzgibbon, Cheree, Young, Samuel N., Hennessy, Patrick J., Smith, Phoebe P. C., Webb, Andrew I., Czabotar, Peter E., Koide, Shohei, and Murphy, James M.
- Published
- 2020
8. The regulation of necroptosis by post-translational modifications
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Meng, Yanxiang, Sandow, Jarrod J., Czabotar, Peter E., and Murphy, James M.
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- 2021
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9. Clinical MDR1 inhibitors enhance Smac-mimetic bioavailability to kill murine LSCs and improve survival in AML models
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Morrish, Emma, Copeland, Anthony, Moujalled, Donia M., Powell, Jason A., Silke, Natasha, Lin, Ann, Jarman, Kate E., Sandow, Jarrod J., Ebert, Gregor, Mackiewicz, Liana, Beach, Jessica A., Christie, Elizabeth L., Lewis, Alexander C., Pomilio, Giovanna, Fischer, Karla C., MacPherson, Laura, Bowtell, David D.L., Webb, Andrew I., Pellegrini, Marc, Dawson, Mark A., Pitson, Stuart M., Wei, Andrew H., Silke, John, and Brumatti, Gabriela
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- 2020
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10. Phosphorylation by Aurora B kinase regulates caspase-2 activity and function
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Lim, Yoon, De Bellis, Dylan, Sandow, Jarrod J., Capalbo, Luisa, D’Avino, Pier Paolo, Murphy, James M., Webb, Andrew I., Dorstyn, Loretta, and Kumar, Sharad
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- 2021
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11. Autophagy captures the retromer-TBC1D5 complex to inhibit receptor recycling.
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Carosi, Julian M., Hein, Leanne K., Sandow, Jarrod J., Dang, Linh V. P., Hattersley, Kathryn, Denton, Donna, Kumar, Sharad, and Sargeant, Timothy J.
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GTPASE-activating protein ,AUTOPHAGY ,AMYLOID beta-protein precursor ,PHYSIOLOGIC salines ,RIBOSOMAL proteins ,PROTEIN kinases ,TUBULINS - Abstract
Retromer prevents the destruction of numerous receptors by recycling them from endosomes to the trans-Golgi network or plasma membrane. This enables retromer to fine-tune the activity of many signaling pathways in parallel. However, the mechanism(s) by which retromer function adapts to environmental fluctuations such as nutrient withdrawal and how this affects the fate of its cargoes remains incompletely understood. Here, we reveal that macroautophagy/autophagy inhibition by MTORC1 controls the abundance of retromer
+ endosomes under nutrient-replete conditions. Autophagy activation by chemical inhibition of MTOR or nutrient withdrawal does not affect retromer assembly or its interaction with the RAB7 GAP protein TBC1D5, but rather targets these endosomes for bulk destruction following their capture by phagophores. This process appears to be distinct from amphisome formation. TBC1D5 and its ability to bind to retromer, but not its C-terminal LC3-interacting region (LIR) or nutrient-regulated dephosphorylation, is critical for retromer to be captured by autophagosomes following MTOR inhibition. Consequently, endosomal recycling of its cargoes to the plasma membrane and trans-Golgi network is impaired, leading to their lysosomal turnover. These findings demonstrate a mechanistic link connecting nutrient abundance to receptor homeostasis. Abbreviations: AMPK, 5'-AMP-activated protein kinase; APP, amyloid beta precursor protein; ATG, autophagy related; BafA, bafilomycin A1 ; CQ, chloroquine; DMEM, Dulbecco's minimum essential medium; DPBS, Dulbecco's phosphate-buffered saline; EBSS, Earle's balanced salt solution; FBS, fetal bovine serum; GAP, GTPase-activating protein; MAP1LC3/LC3, microtubule associated protein 1 light chain 3; LIR, LC3-interacting region; LANDO, LC3-associated endocytosis; LP, leupeptin and pepstatin; MTOR, mechanistic target of rapamycin kinase; MTORC1, MTOR complex 1; nutrient stress, withdrawal of amino acids and serum; PDZ, DLG4/PSD95, DLG1, and TJP1/zo-1; RPS6, ribosomal protein S6; RPS6KB1/S6K1, ribosomal protein S6 kinase B1; SLC2A1/GLUT1, solute carrier family 2 member 1; SORL1, sortillin related receptor 1; SORT1, sortillin 1; SNX, sorting nexin; TBC1D5, TBC1 domain family member 5; ULK1, unc-51 like autophagy activating kinase 1; WASH, WASH complex subunit. [ABSTRACT FROM AUTHOR]- Published
- 2024
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12. Conformational interconversion of MLKL and disengagement from RIPK3 precede cell death by necroptosis
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Garnish, Sarah E., Meng, Yanxiang, Koide, Akiko, Sandow, Jarrod J., Denbaum, Eric, Jacobsen, Annette V., Yeung, Wayland, Samson, Andre L., Horne, Christopher R., Fitzgibbon, Cheree, Young, Samuel N., Smith, Phoebe P. C., Webb, Andrew I., Petrie, Emma J., Hildebrand, Joanne M., Kannan, Natarajan, Czabotar, Peter E., Koide, Shohei, and Murphy, James M.
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- 2021
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13. Granulovirus PK-1 kinase activity relies on a side-to-side dimerization mode centered on the regulatory αC helix
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Oliver, Michael R., Horne, Christopher R., Shrestha, Safal, Keown, Jeremy R., Liang, Lung-Yu, Young, Samuel N., Sandow, Jarrod J., Webb, Andrew I., Goldstone, David C., Lucet, Isabelle S., Kannan, Natarajan, Metcalf, Peter, and Murphy, James M.
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- 2021
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14. Human RIPK3 maintains MLKL in an inactive conformation prior to cell death by necroptosis
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Meng, Yanxiang, Davies, Katherine A., Fitzgibbon, Cheree, Young, Samuel N., Garnish, Sarah E., Horne, Christopher R., Luo, Cindy, Garnier, Jean-Marc, Liang, Lung-Yu, Cowan, Angus D., Samson, Andre L., Lessene, Guillaume, Sandow, Jarrod J., Czabotar, Peter E., and Murphy, James M.
- Published
- 2021
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15. Identification of novel interacting partners of the NEDD4 ubiquitin ligase in mouse testis
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Manning, JantinaA., Windley, Simon P., Sandow, Jarrod J., Shah, Sonia S., Western, Patrick, Wilhelm, Dagmar, and Kumar, Sharad
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- 2020
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16. BAK core dimers bind lipids and can be bridged by them
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Cowan, Angus D., Smith, Nicholas A., Sandow, Jarrod J., Kapp, Eugene A., Rustam, Yepy H., Murphy, James M., Brouwer, Jason M., Bernardini, Jonathan P., Roy, Michael J., Wardak, Ahmad Z., Tan, Iris K., Webb, Andrew I., Gulbis, Jacqueline M., Smith, Brian J., Reid, Gavin E., Dewson, Grant, Colman, Peter M., and Czabotar, Peter E.
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- 2020
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17. A small molecule interacts with VDAC2 to block mouse BAK-driven apoptosis
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van Delft, Mark F., Chappaz, Stephane, Khakham, Yelena, Bui, Chinh T., Debrincat, Marlyse A., Lowes, Kym N., Brouwer, Jason M., Grohmann, Christoph, Sharp, Phillip P., Dagley, Laura F., Li, Lucy, McArthur, Kate, Luo, Meng-Xiao, Chin, Hui San, Fairlie, W. Douglas, Lee, Erinna F., Segal, David, Duflocq, Stephane, Lessene, Romina, Bernard, Sabrina, Peilleron, Laure, Nguyen, Thao, Miles, Caroline, Wan, Soo San, Lane, Rachael M., Wardak, Ahmad, Lackovic, Kurt, Colman, Peter M., Sandow, Jarrod J., Webb, Andrew I., Czabotar, Peter E., Dewson, Grant, Watson, Keith G., Huang, David C. S., Lessene, Guillaume, and Kile, Benjamin T.
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- 2019
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18. Structure of Plasmodium falciparum Rh5–CyRPA–Ripr invasion complex
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Wong, Wilson, Huang, Rick, Menant, Sebastien, Hong, Chuan, Sandow, Jarrod J., Birkinshaw, Richard W., Healer, Julie, Hodder, Anthony N., Kanjee, Usheer, Tonkin, Christopher J., Heckmann, Denise, Soroka, Vladislav, Søgaard, Teit Max Moscote, Jørgensen, Thomas, Duraisingh, Manoj T., Czabotar, Peter E., de Jongh, Willem A., Tham, Wai-Hong, Webb, Andrew I., Yu, Zhiheng, and Cowman, Alan F.
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- 2019
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19. Plasmepsin V cleaves malaria effector proteins in a distinct endoplasmic reticulum translocation interactome for export to the erythrocyte
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Marapana, Danushka S., Dagley, Laura F., Sandow, Jarrod J., Nebl, Thomas, Triglia, Tony, Pasternak, Michał, Dickerman, Benjamin K., Crabb, Brendan S., Gilson, Paul R., Webb, Andrew I., Boddey, Justin A., and Cowman, Alan F.
- Published
- 2018
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20. Distinct pseudokinase domain conformations underlie divergent activation mechanisms among vertebrate MLKL orthologues
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Davies, Katherine A., Fitzgibbon, Cheree, Young, Samuel N., Garnish, Sarah E., Yeung, Wayland, Coursier, Diane, Birkinshaw, Richard W., Sandow, Jarrod J., Lehmann, Wil I. L., Liang, Lung-Yu, Lucet, Isabelle S., Chalmers, James D., Patrick, Wayne M., Kannan, Natarajan, Petrie, Emma J., Czabotar, Peter E., and Murphy, James M.
- Published
- 2020
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21. Necroptotic signaling is primed in Mycobacterium tuberculosis-infected macrophages, but its pathophysiological consequence in disease is restricted
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Stutz, Michael D., Ojaimi, Samar, Allison, Cody, Preston, Simon, Arandjelovic, Philip, Hildebrand, Joanne M., Sandow, Jarrod J., Webb, Andrew I., Silke, John, Alexander, Warren S., and Pellegrini, Marc
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- 2018
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22. A combined immunopeptidomics, proteomics, and cell surface proteomics approach to identify immunotherapy targets for diffuse intrinsic pontine glioma.
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Pandey, Kirti, Wang, Stacie S., Mifsud, Nicole A., Faridi, Pouya, Davenport, Alexander J., Webb, Andrew I., Sandow, Jarrod J., Ayala, Rochelle, Monje, Michelle, Cross, Ryan S., Ramarathinam, Sri H., Jenkins, Misty R., and Purcell, Anthony W.
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PROTEOMICS ,CELL surface antigens ,EPIDERMAL growth factor receptors ,EPHRIN receptors ,GLIOMAS ,GROWTH factors - Abstract
Introduction: Diffuse intrinsic pontine glioma (DIPG), recently reclassified as a subtype of diffuse midline glioma, is a highly aggressive brainstem tumor affecting children and young adults, with no cure and a median survival of only 9 months. Conventional treatments are ineffective, highlighting the need for alternative therapeutic strategies such as cellular immunotherapy. However, identifying unique and tumor-specific cell surface antigens to target with chimeric antigen receptor (CAR) or T-cell receptor (TCR) therapies is challenging. Methods: In this study, a multi-omics approach was used to interrogate patientderived DIPG cell lines and to identify potential targets for immunotherapy. Results: Through immunopeptidomics, a range of targetable peptide antigens fromcancer testis and tumor-associated antigens as well as peptides derived from human endogenous retroviral elements were identified. Proteomics analysis also revealed upregulation of potential drug targets and cell surface proteins such as Cluster of differentiation 27 (CD276) B7 homolog 3 protein (B7H3), Interleukin 13 alpha receptor 2 (IL-13Ra2), Human Epidermal Growth Factor Receptor 3 (HER2), Ephrin Type-A Receptor 2 (EphA2), and Ephrin Type-A Receptor 3 (EphA3). Discussion: The results of this study provide a valuable resource for the scientific community to accelerate immunotherapeutic approaches for DIPG. Identifying potential targets for CAR and TCR therapies could open up new avenues for treating this devastating disease. [ABSTRACT FROM AUTHOR]
- Published
- 2023
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23. VDAC2 enables BAX to mediate apoptosis and limit tumor development
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Chin, Hui San, Li, Mark X., Tan, Iris K. L., Ninnis, Robert L., Reljic, Boris, Scicluna, Kristen, Dagley, Laura F., Sandow, Jarrod J., Kelly, Gemma L., Samson, Andre L., Chappaz, Stephane, Khaw, Seong L., Chang, Catherine, Morokoff, Andrew, Brinkmann, Kerstin, Webb, Andrew, Hockings, Colin, Hall, Cathrine M., Kueh, Andrew J., Ryan, Michael T., Kluck, Ruth M., Bouillet, Philippe, Herold, Marco J., Gray, Daniel H. D., Huang, David C. S., van Delft, Mark F., and Dewson, Grant
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- 2018
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24. Signalling by the βc family of cytokines
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Hercus, Timothy R., Dhagat, Urmi, Kan, Winnie L.T., Broughton, Sophie E., Nero, Tracy L., Perugini, Michelle, Sandow, Jarrod J., D’Andrea, Richard J., Ekert, Paul G., Hughes, Timothy, Parker, Michael W., and Lopez, Angel F.
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- 2013
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25. CD98 defines a metabolically flexible, proinflammatory subset of low‐density neutrophils in systemic lupus erythematosus.
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Martin, Katherine R., Day, Jessica A., Hansen, Jacinta A., D'Silva, Damian B., Wong, Huon L., Garnham, Alexandra, Sandow, Jarrod J., Nijagal, Brunda, Wilson, Nicholas, and Wicks, Ian P.
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SYSTEMIC lupus erythematosus ,GRANULOCYTE-colony stimulating factor ,NEUTROPHILS ,ESSENTIAL amino acids ,MEMBRANE proteins ,ADENOSINE triphosphate ,GRANULOCYTES - Abstract
Background: Low‐density neutrophils (LDN) are a distinct subset of neutrophils rarely detected in healthy people but appear in the blood of patients with autoimmune diseases, including systemic lupus erythematosus (SLE), and are mobilised in response to granulocyte colony‐stimulating factor (G‐CSF). The aim of this study was to identify novel mechanisms responsible for the pathogenic capacity of LDN in SLE. Methods: Neutrophils were isolated from donors treated with G‐CSF, and whole‐cell proteomic analysis was performed on LDN and normal‐density neutrophils. Results: CD98 is significantly upregulated in LDN from G‐CSF donors and defines a subset of LDN within the blood of SLE patients. CD98 is a transmembrane protein that dimerises with L‐type amino acid transporters. We show that CD98 is responsible for the increased bioenergetic capacity of LDN. CD98 on LDN mediates the uptake of essential amino acids that are used by mitochondria to produce adenosine triphosphate, especially in the absence of glucose. Inhibition of CD98 reduces the metabolic flexibility of this population, which may limit their pathogenic capacity. CD98+ LDN produce more proinflammatory cytokines and chemokines than their normal density counterparts and are resistant to apoptosis, which may also contribute to tissue inflammation and end organ damage in SLE. Conclusions: CD98 provides a phenotypic marker for LDN that facilitates identification of this population without density‐gradient separation and represents a novel therapeutic target to limit its pathogenic capacity. [ABSTRACT FROM AUTHOR]
- Published
- 2023
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26. Tankyrase-mediated ADP-ribosylation is a regulator of TNF-induced death.
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Lin Liu, Sandow, Jarrod J., Pedrioli, Deena M. Leslie, Samson, Andre L., Silke, Natasha, Kratina, Tobias, Ambrose, Rebecca L., Doerflinger, Marcel, Zhaoqing Hu, Morrish, Emma, Diep Chau, Kueh, Andrew J., Fitzibbon, Cheree, Pellegrini, Marc, Pearson, Jaclyn S., Hottiger, Michael O., Webb, Andrew I., Lalaoui, Najoua, and Silke, John
- Subjects
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CELL death , *DEATH receptors , *DOXYCYCLINE , *LIFE sciences , *ADP-ribosylation , *SARS-CoV-2 , *GREEN fluorescent protein - Abstract
The article presents a study which explores that Tankyrase-mediated ADP-ribosylation, a regulator of Tumor necrosis factor (TNF)-induced death . It mentions that findings of the study suggests that disruption of ADP-ribosylation during an infection can prime a cell to retaliate with an inflammatory cell death.
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- 2022
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27. K‐29 linked ubiquitination of Arrdc4 regulates its function in extracellular vesicle biogenesis.
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Farooq, Ammara Usman, Gembus, Kelly, Sandow, Jarrod J., Webb, Andrew, Mathivanan, Suresh, Manning, Jantina A., Shah, Sonia S., Foot, Natalie J., and Kumar, Sharad
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EXTRACELLULAR vesicles ,UBIQUITINATION ,UBIQUITIN ligases ,CELL communication ,ARRESTINS - Abstract
Extracellular vesicles (EVs) are important mediators of intercellular communication. However, EV biogenesis remains poorly understood. We previously defined a role for Arrdc4 (Arrestin domain containing protein 4), an adaptor for Nedd4 family ubiquitin ligases, in the biogenesis of EVs. Here we report that ubiquitination of Arrdc4 is critical for its role in EV secretion. We identified five potential ubiquitinated lysine residues in Arrdc4 using mass spectrometry. By analysing Arrdc4 lysine mutants we discovered that lysine 270 (K270) is critical for Arrdc4 function in EV biogenesis. Arrdc4K270R mutation caused a decrease in the number of EVs released by cells compared to Arrdc4WT, and a reduction in trafficking of divalent metal transporter (DMT1) into EVs. Furthermore, we also observed a decrease in DMT1 activity and an increase in its intracellular degradation in the presence of Arrdc4K270R. K270 was found to be ubiquitinated with K‐29 polyubiquitin chains by the ubiquitin ligase Nedd4‐2. Thus, our results uncover a novel role of K‐29 polyubiquitin chains in Arrdc4‐mediated EV biogenesis and protein trafficking. [ABSTRACT FROM AUTHOR]
- Published
- 2022
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28. Dynamic reconfiguration of pro‐apoptotic BAK on membranes.
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Sandow, Jarrod J, Tan, Iris KL, Huang, Alan S, Masaldan, Shashank, Bernardini, Jonathan P, Wardak, Ahmad Z, Birkinshaw, Richard W, Ninnis, Robert L, Liu, Ziyan, Dalseno, Destiny, Lio, Daisy, Infusini, Giuseppi, Czabotar, Peter E, Webb, Andrew I, and Dewson, Grant
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CELL death , *HYDROGEN-deuterium exchange , *LIPOSOMES , *MASS spectrometry , *CYTOCHROME c , *SITE-specific mutagenesis - Abstract
BAK and BAX, the effectors of intrinsic apoptosis, each undergo major reconfiguration to an activated conformer that self‐associates to damage mitochondria and cause cell death. However, the dynamic structural mechanisms of this reconfiguration in the presence of a membrane have yet to be fully elucidated. To explore the metamorphosis of membrane‐bound BAK, we employed hydrogen‐deuterium exchange mass spectrometry (HDX‐MS). The HDX‐MS profile of BAK on liposomes comprising mitochondrial lipids was consistent with known solution structures of inactive BAK. Following activation, HDX‐MS resolved major reconfigurations in BAK. Mutagenesis guided by our HDX‐MS profiling revealed that the BCL‐2 homology (BH) 4 domain maintains the inactive conformation of BAK, and disrupting this domain is sufficient for constitutive BAK activation. Moreover, the entire N‐terminal region preceding the BAK oligomerisation domains became disordered post‐activation and remained disordered in the activated oligomer. Removal of the disordered N‐terminus did not impair, but rather slightly potentiated, BAK‐mediated membrane permeabilisation of liposomes and mitochondria. Together, our HDX‐MS analyses reveal new insights into the dynamic nature of BAK activation on a membrane, which may provide new opportunities for therapeutic targeting. SYNOPSIS: While BAK is a known executioner protein in apoptotic cell death, how it changes conformation to form oligomeric mitochondrial pores to kill cells remains unclear. Hydrogen‐deuterium exchange mass spectrometry and site‐directed mutagenesis now resolve key events in BAK activation as it occurs on membranes and mitochondria. Hydrogen‐deuterium exchange mass spectrometry provides new insight into conformation changes of BAK on a mitochondria‐like membrane.The BAK BH4 domain constrains BAK activity in the absence of a death stimulus.During BAK activation and oligomerisation, amino acids preceding the BH3 domain become exposed and disordered.The disordered N‐terminus is not required for BAK to permeabilise membranes and mediate cytochrome c release and may rather inhibit it. [ABSTRACT FROM AUTHOR]
- Published
- 2021
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29. Cp1/cathepsin L is required for autolysosomal clearance in Drosophila.
- Author
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Xu, Tianqi, Nicolson, Shannon, Sandow, Jarrod J., Dayan, Sonia, Jiang, Xin, Manning, Jantina A., Webb, Andrew I., Kumar, Sharad, and Denton, Donna
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GREEN fluorescent protein ,CATHEPSIN B ,CELL survival ,PEPTIDASE ,DROSOPHILA ,CATHEPSIN D ,MICROPHTHALMIA-associated transcription factor - Abstract
Macroautophagy/autophagy is a highly conserved lysosomal degradative pathway important for maintaining cellular homeostasis. Much of our current knowledge of autophagy is focused on the initiation steps in this process. Recently, an understanding of later steps, particularly lysosomal fusion leading to autolysosome formation and the subsequent role of lysosomal enzymes in degradation and recycling, is becoming evident. Autophagy can function in both cell survival and cell death, however, the mechanisms that distinguish adaptive/survival autophagy from autophagy-dependent cell death remain to be established. Here, using proteomic analysis of Drosophila larval midguts during degradation, we identify a group of proteins with peptidase activity, suggesting a role in autophagy-dependent cell death. We show that Cp1/cathepsin L-deficient larval midgut cells accumulate aberrant autophagic vesicles due to a block in autophagic flux, yet later stages of midgut degradation are not compromised. The accumulation of large aberrant autolysosomes in the absence of Cp1 appears to be the consequence of decreased degradative capacity as they contain undigested cytoplasmic material, rather than a defect in autophagosome-lysosome fusion. Finally, we find that other cathepsins may also contribute to proper autolysosomal degradation in Drosophila larval midgut cells. Our findings provide evidence that cathepsins play an essential role in the autolysosome to maintain basal autophagy flux by balancing autophagosome production and turnover. Abbreviations: 26-29-p: 26-29kD-proteinase; ADCD: autophagy-dependent cell death; Atg8a: Autophagy-related protein 8a; Cp1/cathepsin L: Cysteine proteinase-1; CtsB: Cathepsin B; cathD: cathepsin D; CtsF: Cathepsin F; GFP: green fluorescent protein; LAMP1: lysosomal-associated membrane protein 1; Mitf: microphthalmia associated transcription factor; PCA: principal component analysis; RNAi: RNA interference; RPF: relative to puparium formation [ABSTRACT FROM AUTHOR]
- Published
- 2021
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30. Proteomic analyses reveal that immune integrins are major targets for regulation by Membrane‐Associated Ring‐CH (MARCH) proteins MARCH2, 3, 4 and 9.
- Author
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Sandow, Jarrod J., Webb, Andrew I., Stockwell, Dina, Kershaw, Nadia J., Tan, Cyrus, Ishido, Satoshi, Alexander, Warren S., Hilton, Douglas J., Babon, Jeffrey J., and Nicola, Nicos A.
- Published
- 2021
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31. Arrdc4‐dependent extracellular vesicle biogenesis is required for sperm maturation.
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Foot, Natalie J., Gonzalez, Macarena B., Gembus, Kelly, Fonseka, Pamali, Sandow, Jarrod J., Nguyen, Thuy Tien, Tran, Diana, Webb, Andrew I., Mathivanan, Suresh, Robker, Rebecca L., and Kumar, Sharad
- Subjects
EXTRACELLULAR vesicles ,MALE reproductive organs ,ACROSOME reaction ,GENITALIA ,ZONA pellucida - Abstract
Extracellular vesicles (EVs) are important players in cell to cell communication in reproductive systems. Notably, EVs have been found and characterized in the male reproductive tract, however, direct functional evidence for their importance in mediating sperm function is lacking. We have previously demonstrated that Arrdc4, a member of the α‐arrestin protein family, is involved in extracellular vesicle biogenesis and release. Here we show that Arrdc4‐mediated extracellular vesicle biogenesis is required for proper sperm function. Sperm from Arrdc4–/– mice develop normally through the testis but fail to acquire adequate motility and fertilization capabilities through the epididymis, as observed by reduced motility, premature acrosome reaction, reduction in zona pellucida binding and two‐cell embryo production. We found a significant reduction in extracellular vesicle production by Arrdc4–/‐ epididymal epithelial cells, and further, supplementation of Arrdc4–/– sperm with additional vesicles dampened the acrosome reaction defect and restored zona pellucida binding. These results indicate that Arrdc4 is important for proper sperm maturation through the control of extracellular vesicle biogenesis. [ABSTRACT FROM AUTHOR]
- Published
- 2021
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32. The search for RNA-binding proteins: a technical and interdisciplinary challenge.
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Smith, Jeffrey M., Sandow, Jarrod J., and Webb, Andrew I.
- Subjects
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RNA-binding proteins , *RNA-protein interactions , *RNA sequencing , *GENE expression , *PROTEOMICS - Abstract
RNA-binding proteins are customarily regarded as important facilitators of gene expression. In recent years, RNA–protein interactions have also emerged as a pervasive force in the regulation of homeostasis. The compendium of proteins with provable RNA-binding function has swelled from the hundreds to the thousands astride the partnership of mass spectrometry-based proteomics and RNA sequencing. At the foundation of these advances is the adaptation of RNA-centric capture methods that can extract bound protein that has been cross-linked in its native environment. These methods reveal snapshots in time displaying an extensive network of regulation and a wealth of data that can be used for both the discovery of RNA-binding function and the molecular interfaces at which these interactions occur. This review will focus on the impact of these developments on our broader perception of post-transcriptional regulation, and how the technical features of current capture methods, as applied in mammalian systems, create a challenging medium for interpretation by systems biologists and target validation by experimental researchers. [ABSTRACT FROM AUTHOR]
- Published
- 2021
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33. Crystal structure of the hinge domain of Smchd1 reveals its dimerization mode and nucleic acid–binding residues.
- Author
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Chen, Kelan, Birkinshaw, Richard W., Gurzau, Alexandra D., Wanigasuriya, Iromi, Wang, Ruoyun, Iminitoff, Megan, Sandow, Jarrod J., Young, Samuel N., Hennessy, Patrick J., Willson, Tracy A., Heckmann, Denise A., Webb, Andrew I., Blewitt, Marnie E., Czabotar, Peter E., and Murphy, James M.
- Subjects
FACIOSCAPULOHUMERAL muscular dystrophy ,CRYSTAL structure ,DIMERIZATION ,HINGES ,MUSCULAR dystrophy ,NUCLEIC acids - Abstract
Illuminating dimerization: Proteins of the SMC family are chromosomal organizers involved in sister chromatid cohesion, chromosome condensation, and DNA repair. Unlike other eukaryotic family members, SMCHD1 forms homodimers, rather than heterodimers, and has a distinct domain architecture. Dysregulation of SMCHD1 function results in a form of muscular dystrophy and a developmental disorder. Chen et al. solved the x-ray crystal structure of the Smchd1 hinge domain, which is important for homodimerization and nucleic acid binding. Site-directed mutagenesis studies identified critical residues involved in SMCHD1 function in cells. Together, these data suggest how mutations in the SMCHD1 hinge domain contribute to human disease. Structural maintenance of chromosomes flexible hinge domain containing 1 (SMCHD1) is an epigenetic regulator in which polymorphisms cause the human developmental disorder, Bosma arhinia micropthalmia syndrome, and the degenerative disease, facioscapulohumeral muscular dystrophy. SMCHD1 is considered a noncanonical SMC family member because its hinge domain is C-terminal, because it homodimerizes rather than heterodimerizes, and because SMCHD1 contains a GHKL-type, rather than an ABC-type ATPase domain at its N terminus. The hinge domain has been previously implicated in chromatin association; however, the underlying mechanism involved and the basis for SMCHD1 homodimerization are unclear. Here, we used x-ray crystallography to solve the three-dimensional structure of the Smchd1 hinge domain. Together with structure-guided mutagenesis, we defined structural features of the hinge domain that participated in homodimerization and nucleic acid binding, and we identified a functional hotspot required for chromatin localization in cells. This structure provides a template for interpreting the mechanism by which patient polymorphisms within the SMCHD1 hinge domain could compromise function and lead to facioscapulohumeral muscular dystrophy. [ABSTRACT FROM AUTHOR]
- Published
- 2020
- Full Text
- View/download PDF
34. A class of γδ T cell receptors recognize the underside of the antigen-presenting molecule MR1.
- Author
-
Le Nours, Jérôme, Gherardin, Nicholas A., Ramarathinam, Sri H., Awad, Wael, Wiede, Florian, Gully, Benjamin S., Khandokar, Yogesh, Praveena, T., Wubben, Jacinta M., Sandow, Jarrod J., Webb, Andrew I., von Borstel, Anouk, Rice, Michael T., Redmond, Samuel J., Seneviratna, Rebecca, Sandoval-Romero, Maria L., Shihan Li, Souter, Michael N. T., Eckle, Sidonia B. G., and Corbett, Alexandra J.
- Published
- 2019
- Full Text
- View/download PDF
35. Universal Solid-Phase Protein Preparation (USP3) for Bottom-up and Top-down Proteomics.
- Author
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Dagley, Laura F., Infusini, Giuseppe, Larsen, Rune H., Sandow, Jarrod J., and Webb, Andrew I.
- Published
- 2019
- Full Text
- View/download PDF
36. Conformational switching of the pseudokinase domain promotes human MLKL tetramerization and cell death by necroptosis.
- Author
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Petrie, Emma J., Sandow, Jarrod J., Jacobsen, Annette V., Smith, Brian J., Griffin, Michael D. W., Lucet, Isabelle S., Weiwen Dai, Young, Samuel N., Tanzer, Maria C., Wardak, Ahmad, Lung-Yu Liang, Cowan, Angus D., Hildebrand, Joanne M., Kersten, Wilhelmus J. A., Lessene, Guillaume, Silke, John, Czabotar, Peter E., Webb, Andrew I., and Murphy, James M.
- Subjects
CELL death ,MOLECULAR switches ,CELL membranes - Abstract
Necroptotic cell death is mediated by the most terminal known effector of the pathway, MLKL. Precisely how phosphorylation of the MLKL pseudokinase domain activation loop by the upstream kinase, RIPK3, induces unmasking of the N-terminal executioner four-helix bundle (4HB) domain of MLKL, higher-order assemblies, and permeabilization of plasma membranes remains poorly understood. Here, we reveal the existence of a basal monomeric MLKL conformer present in human cells prior to exposure to a necroptotic stimulus. Following activation, toggling within the MLKL pseudokinase domain promotes 4HB domain disengagement from the pseudokinase domain αC helix and pseudocatalytic loop, to enable formation of a necroptosis-inducing tetramer. In contrast to mouse MLKL, substitution of RIPK3 substrate sites in the human MLKL pseudokinase domain completely abrogated necroptotic signaling. Therefore, while the pseudokinase domains of mouse and human MLKL function as molecular switches to control MLKL activation, the underlying mechanism differs between species. [ABSTRACT FROM AUTHOR]
- Published
- 2018
- Full Text
- View/download PDF
37. Discovery and Validation of Novel Protein Biomarkers in Ovarian Cancer Patient Urine.
- Author
-
Sandow, Jarrod J., Rainczuk, Adam, Infusini, Giuseppe, Makanji, Ming, Bilandzic, Maree, Wilson, Amy L., Fairweather, Nicole, Stanton, Peter G., Garama, Daniel, Gough, Daniel, Jobling, Thomas W., Webb, Andrew I., and Stephens, Andrew N.
- Published
- 2018
- Full Text
- View/download PDF
38. Quantitative proteomic analysis of EZH2 inhibition in acute myeloid leukemia reveals the targets and pathways that precede the induction of cell death.
- Author
-
Sandow, Jarrod J., Infusini, Giuseppe, Holik, Aliaksei Z., Brumatti, Gabriela, Averink, Tessa V., Ekert, Paul G., and Webb, Andrew I.
- Published
- 2017
- Full Text
- View/download PDF
39. Enzymatic Characterization of Wild-Type and Mutant Janus Kinase 1.
- Author
-
Liau, Nicholas P. D., Laktyushin, Artem, Morris, Rhiannon, Sandow, Jarrod J., Nicola, Nicos A., Kershaw, Nadia J., and Babon, Jeffrey J.
- Subjects
PROTEIN metabolism ,ENZYME metabolism ,CELL receptors ,CYTOKINES ,LEUKEMIA ,LYMPHOMAS ,GENETIC mutation ,MYELOPROLIFERATIVE neoplasms ,PHOSPHOTRANSFERASES ,POLYCYTHEMIA vera ,PHENOTYPES ,IN vitro studies ,JANUS kinases - Abstract
Janus kinases (JAKs) are found constitutively associated with cytokine receptors and are present in an inactive state prior to cytokine exposure. Activating mutations of JAKs are causative for a number of leukemias, lymphomas, and myeloproliferative diseases. In particular, the JAK2
V617F mutant is found in most human cases of polycythemia vera, a disease characterized by over-production of erythrocytes. The V617F mutation is found in the pseudokinase domain of JAK2 and it leads to cytokine-independent activation of the kinase, as does the orthologous mutation in other JAK-family members. The mechanism whereby this mutation hyperactivates these kinases is not well understood, primarily due to the fact that the full-length JAK proteins are difficult to produce for structural and kinetic studies. Here we have overcome this limitation to perform a series of enzymatic analyses on full-length JAK1 and its constitutively active mutant form (JAK1V658F ). Consistent with previous studies, we show that the presence of the pseudokinase domain leads to a dramatic decrease in enzymatic activity with no further decrease from the presence of the FERM or SH2 domains. However, we find that the mutant kinase, in vitro, is indistinguishable from the wild-type enzyme in every measurable parameter tested: KM (ATP), KM (substrate), kcat , receptor binding, thermal stability, activation rate, dephosphorylation rate, and inhibitor affinity. These results show that the V658F mutation does not enhance the intrinsic enzymatic activity of JAK. Rather this data is more consistent with a model in which there are cellular processes and interactions that prevent JAK from being activated in the absence of cytokine and it is these constraints that are affected by disease-causing mutations. [ABSTRACT FROM AUTHOR]- Published
- 2019
- Full Text
- View/download PDF
40. Viral MLKL Homologs Subvert Necroptotic Cell Death by Sequestering Cellular RIPK3.
- Author
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Petrie, Emma J., Sandow, Jarrod J., Lehmann, Wil I.L., Liang, Lung-Yu, Coursier, Diane, Young, Samuel N., Kersten, Wilhelmus J.A., Fitzgibbon, Cheree, Samson, André L., Jacobsen, Annette V., Lowes, Kym N., Au, Amanda E., Jousset Sabroux, Hélène, Lalaoui, Najoua, Webb, Andrew I., Lessene, Guillaume, Manning, Gerard, Lucet, Isabelle S., and Murphy, James M.
- Abstract
Necroptotic cell death has been implicated in many human pathologies and is thought to have evolved as an innate immunity mechanism. The pathway relies on two key effectors: the kinase receptor-interacting protein kinase 3 (RIPK3) and the terminal effector, the pseudokinase mixed-lineage kinase-domain-like (MLKL). We identify proteins with high sequence similarity to the pseudokinase domain of MLKL in poxvirus genomes. Expression of these proteins from the BeAn 58058 and Cotia poxviruses, but not swinepox, in human and mouse cells blocks cellular MLKL activation and necroptotic cell death. We show that viral MLKL-like proteins function as dominant-negative mimics of host MLKL, which inhibit necroptosis by sequestering RIPK3 via its kinase domain to thwart MLKL engagement and phosphorylation. These data support an ancestral role for necroptosis in defense against pathogens. Furthermore, mimicry of a cellular pseudokinase by a pathogen adds to the growing repertoire of functions performed by pseudokinases in signal transduction. • Some poxviruses encode proteins with homology to mammalian MLKL pseudokinase domains • BAV and Cotia viral MLKL proteins inhibit necroptotic death of human and mouse cells • These viral MLKL proteins target RIPK3 to block activation of cellular MLKL • Viral MLKL proteins inhibit RIPK3 via species-specific mechanisms Petrie et al. identify proteins encoded by some poxviruses with homology to the pseudokinase domain of the terminal necroptosis effector, MLKL. Via species-specific mechanisms, viral MLKL proteins block necroptotic death in human and mouse cells by sequestering RIPK3 to prevent phosphorylation and activation of MLKL. [ABSTRACT FROM AUTHOR]
- Published
- 2019
- Full Text
- View/download PDF
41. Parkin inhibits BAK and BAX apoptotic function by distinct mechanisms during mitophagy.
- Author
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Bernardini, Jonathan P, Brouwer, Jason M, Tan, Iris KL, Sandow, Jarrod J, Huang, Shuai, Stafford, Che A, Bankovacki, Aleksandra, Riffkin, Christopher D, Wardak, Ahmad Z, Czabotar, Peter E, Lazarou, Michael, and Dewson, Grant
- Subjects
PARKIN (Protein) ,UBIQUITIN ligases ,MITOCHONDRIA ,UBIQUITINATION ,APOPTOSIS - Abstract
The E3 ubiquitin ligase Parkin is a key effector of the removal of damaged mitochondria by mitophagy. Parkin determines cell fate in response to mitochondrial damage, with its loss promoting early onset Parkinson's disease and potentially also cancer progression. Controlling a cell's apoptotic response is essential to co‐ordinate the removal of damaged mitochondria. We report that following mitochondrial damage‐induced mitophagy, Parkin directly ubiquitinates the apoptotic effector protein BAK at a conserved lysine in its hydrophobic groove, a region that is crucial for BAK activation by BH3‐only proteins and its homo‐dimerisation during apoptosis. Ubiquitination inhibited BAK activity by impairing its activation and the formation of lethal BAK oligomers. Parkin also suppresses BAX‐mediated apoptosis, but in the absence of BAX ubiquitination suggesting an indirect mechanism. In addition, we find that BAK‐dependent mitochondrial outer membrane permeabilisation during apoptosis promotes PINK1‐dependent Parkin activation. Hence, we propose that Parkin directly inhibits BAK to suppress errant apoptosis, thereby allowing the effective clearance of damaged mitochondria, but also promotes clearance of apoptotic mitochondria to limit their potential pro‐inflammatory effect. Synopsis: The ubiquitin ligase Parkin plays a protective role in neurodegenerative disease by removing damaged mitochondria and preventing apoptosis, and by limiting the functions of pro‐apoptotic effector proteins BAK and BAX. Defective control of apoptosis may contribute to the pathogenesis of early onset Parkinson's Disease caused by Parkin mutations. Parkin activity is induced by mitochondrial damage during apoptosis.Parkin mono‐ and di‐ubiquitinates BAK at a conserved lysine in its hydrophobic groove.Ubiquitination reduces BAK oligomerisation and apoptotic activity on mitochondria.Parkin prevents BAX mitochondrial localisation and apoptotic activity independent of ubiquitination.Certain Parkinson's Disease‐associated Parkin mutants cannot ubiquitinate BAK and restrain it on mitochondria. Autophagic clearance of damaged mitochondria and suppression of apoptotic effector proteins are coordinated functions of the Parkin ubiquitin ligase that are lost upon Parkinson's Disease‐associated mutations. [ABSTRACT FROM AUTHOR]
- Published
- 2019
- Full Text
- View/download PDF
42. A RIPK2 inhibitor delays NOD signalling events yet prevents inflammatory cytokine production.
- Author
-
Nachbur, Ueli, Stafford, Che A., Bankovacki, Aleksandra, Zhan, Yifan, Lindqvist, Lisa M., Fiil, Berthe K., Khakham, Yelena, Ko, Hyun-Ja, Sandow, Jarrod J., Falk, Hendrik, Holien, Jessica K., Chau, Diep, Hildebrand, Joanne, Vince, James E., Sharp, Phillip P., Webb, Andrew I., Jackman, Katherine A., Mühlen, Sabrina, Kennedy, Catherine L., and Lowes, Kym N.
- Published
- 2015
- Full Text
- View/download PDF
43. Role of salt bridges in the dimer interface of 14-3-3ζ in dimer dynamics, N-terminal α-helical order, and molecular chaperone activity.
- Author
-
Woodcock, Joanna M., Goodwin, Katy L., Sandow, Jarrod J., Coolen, Carl, Perugini, Matthew A., Webb, Andrew I., Pitson, Stuart M., Lopez, Angel F., and Carver, John A.
- Subjects
- *
DIMERS , *ADAPTOR proteins , *MOLECULAR chaperones , *HYDROGEN-deuterium exchange , *ULTRACENTRIFUGATION , *MASS spectrometry - Abstract
The 14-3-3 family of intracellular proteins are dimeric, multifunctional adaptor proteins that bind to and regulate the activities of many important signaling proteins. The subunits within 14-3-3 dimers are predicted to be stabilized by salt bridges that are largely conserved across the 14-3-3 protein family and allow the different isoforms to form heterodimers. Here, we have examined the contributions of conserved salt-bridging residues in stabilizing the dimeric state of 14-3-3ζ. Using analytical ultracentrifugation, our results revealed that Asp21 and Glu89 both play key roles in dimer dynamics and contribute to dimer stability. Furthermore, hydrogen-deuterium exchange coupled with mass spectrometry showed that mutation of Asp21 promoted disorder in the N-terminal helices of 14-3-3ζ, suggesting that this residue plays an important role in maintaining structure across the dimer interface. Intriguingly, a D21N 14-3-3ζ mutant exhibited enhanced molecular chaperone ability that prevented amorphous protein aggregation, suggesting a potential role for N-terminal disorder in 14-3-3ζ's poorly understood chaperone action. Taken together, these results imply that disorder in the N-terminal helices of 14-3-3ζ is a consequence of the dimer-monomer dynamics and may play a role in conferring chaperone function to 14-3-3ζ protein. [ABSTRACT FROM AUTHOR]
- Published
- 2018
- Full Text
- View/download PDF
44. Inflammasome sensor NLRP1 disease variant M1184V promotes autoproteolysis and DPP9 complex formation by stabilizing the FIIND domain.
- Author
-
Moecking, Jonas, Laohamonthonkul, Pawat, Meşe, Kubilay, Hagelueken, Gregor, Steiner, Annemarie, Harapas, Cassandra R., Sandow, Jarrod J., Graves, Jonathan D., Masters, Seth L., and Geyer, Matthias
- Subjects
- *
MOLECULAR dynamics , *INFLAMMATORY bowel diseases , *INFLAMMASOMES , *SURFACE plasmon resonance , *PYRIN (Protein) - Abstract
The inflammasome sensor NLRP1 (nucleotide-binding oligomerization domain-like receptor containing a pyrin domain 1) detects a variety of pathogen-derived molecular patterns to induce an inflammatory immune response by triggering pyroptosis and cytokine release. A number of mutations and polymorphisms of NLRP1 are known to cause autoinflammatory diseases, the functional characterization of which contributes to a better understanding of NLRP1 regulation. Here, we assessed the effect of the common NLRP1 variant M1184V, associated with asthma, inflammatory bowel disease, and diabetes, on the protein level. Our size-exclusion chromatography experiments show that M1184V stabilizes the "function-to-find" domain (FIIND) in a monomeric conformation. This effect is independent of autoproteolysis. In addition, molecular dynamics simulations reveal that the methionine residue increases flexibility within the ZU5 domain, whereas valine decreases flexibility, potentially indirectly stabilizing the catalytic triad responsible for autocleavage. By keeping the FIIND domain monomeric, formation of a multimer of full-length NLRP1 is promoted. We found that the stabilizing effect of the valine further leads to improved dipeptidyl peptidase 9 (DPP9)-binding capacities for the FIIND domain as well as the full-length protein as determined by surface plasmon resonance. Moreover, our immuno-precipitation experiments confirmed increased DPP9 binding for the M1184V protein in cells, consistent with improved formation of an autoinhibited complex with DPP9 in activity as-says. Collectively, our study establishes a molecular rationale for the dichotomous involvement of the NLRP1 variant M1184V in autoimmune syndromes. [ABSTRACT FROM AUTHOR]
- Published
- 2022
- Full Text
- View/download PDF
45. The molecular relationships between apoptosis, autophagy and necroptosis.
- Author
-
Lalaoui, Najoua, Lindqvist, Lisa M., Sandow, Jarrod J., and Ekert, Paul G.
- Subjects
- *
MOLECULAR biology , *APOPTOSIS , *AUTOPHAGY , *CELL death , *PATHOLOGICAL physiology , *CASPASES - Abstract
Cells are constantly subjected to a vast range of potentially lethal insults, which may activate specific molecular pathways that have evolved to kill the cell. Cell death pathways are defined partly by their morphology, and more specifically by the molecules that regulate and enact them. As these pathways become more thoroughly characterized, interesting molecular links between them have emerged, some still controversial and others hinting at the physiological and pathophysiological roles these death pathways play. We describe specific molecular programs controlling cell death, with a focus on some of the distinct features of the pathways and the molecular links between them. [ABSTRACT FROM AUTHOR]
- Published
- 2015
- Full Text
- View/download PDF
46. CDK13 cooperates with CDK12 to control global RNA polymerase II processivity.
- Author
-
Zheng Fan, Devlin, Jennifer R., Hogg, Simon J., Doyle, Maria A., Harrison, Paul F., Todorovski, Izabela, Cluse, Leonie A., Knight, Deborah A., Sandow, Jarrod J., Gregory, Gareth, Fox, Andrew, Beilharz, Traude H., Kwiatkowski, Nicholas, Scott, Nichollas E., Vidakovic, Ana Tufegdzic, Kelly, Gavin P., Svejstrup, Jesper Q., Geyer, Matthias, Gray, Nathanael S., and Vervoort, Stephin J.
- Subjects
- *
RNA polymerase II , *STREPTAVIDIN , *RNA polymerases , *RAPAMYCIN , *BIOCHEMISTRY - Abstract
The article discusses a study according to which cyclin-dependent kinases (CDKs)13 cooperates with CDK12 to control global RNA polymerase II processivity. It mentions that single inhibition of CDK12 or CDK13 induced transcriptional responses associated with cellular growth signaling pathways and/or DNA damage, with minimal effects on cell viability.
- Published
- 2020
- Full Text
- View/download PDF
47. Arrdc4‐dependent extracellular vesicle biogenesis is required for sperm maturation
- Author
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Jarrod J. Sandow, Natalie J. Foot, Sharad Kumar, Pamali Fonseka, Thuy Tien Nguyen, Suresh Mathivanan, Diana N. H. Tran, Rebecca L. Robker, Andrew I. Webb, Kelly M. Gembus, Macarena B. Gonzalez, Foot, Natalie J, Gonzalez, Macarena B, Gembus, Kelly, Fonseka, Pamali, Sandow, Jarrod J, Nguyen, Thuy Tien, Tran, Diana, Webb, Andrew I., Mathivanan, Suresh, Robker, Rebecca L, and Kumar, Sharad
- Subjects
0301 basic medicine ,endocrine system ,Histology ,Acrosome reaction ,sperm maturation ,Motility ,03 medical and health sciences ,0302 clinical medicine ,medicine ,ubiquitin ligase adaptors ,Zona pellucida ,reproductive and urinary physiology ,Uncategorized ,fertility ,QH573-671 ,Chemistry ,urogenital system ,arrestin ,Vesicle ,Cell Biology ,Extracellular vesicle ,Epididymis ,biogenesis ,Sperm ,Cell biology ,030104 developmental biology ,medicine.anatomical_structure ,030220 oncology & carcinogenesis ,extracellular vesicles ,Cytology ,Biogenesis - Abstract
Refereed/Peer-reviewed Extracellular vesicles (EVs) are important players in cell to cell communication in reproductive systems. Notably, EVs have been found and characterized in the male reproductive tract, however, direct functional evidence for their importance in mediating sperm function is lacking. We have previously demonstrated that Arrdc4, a member of the α-arrestin protein family, is involved in extracellular vesicle biogenesis and release. Here we show that Arrdc4-mediated extracellular vesicle biogenesis is required for proper sperm function. Sperm from Arrdc4–/– mice develop normally through the testis but fail to acquire adequate motility and fertilization capabilities through the epididymis, as observed by reduced motility, premature acrosome reaction, reduction in zona pellucida binding and two-cell embryo production. We found a significant reduction in extracellular vesicle production by Arrdc4–/- epididymal epithelial cells, and further, supplementation of Arrdc4–/– sperm with additional vesicles dampened the acrosome reaction defect and restored zona pellucida binding. These results indicate that Arrdc4 is important for proper sperm maturation through the control of extracellular vesicle biogenesis. © 2021 The Authors. Journal of Extracellular Vesicles published by Wiley Periodicals, LLC on behalf of the International Society for Extracellular Vesicles.
- Published
- 2021
48. Identification of novel interacting partners of the NEDD4 ubiquitin ligase in mouse testis
- Author
-
Dagmar Wilhelm, Jantina A. Manning, Patrick S Western, Sonia S. Shah, Sharad Kumar, Jarrod J. Sandow, Simon P. Windley, Manning, Jantina A, Windley, Simon P, Sandow, Jarrod J, Shah, Sonia S, Western, Patrick, Wilhelm, Dagmar, and Kumar, Sharad
- Subjects
Male ,Proteomics ,0301 basic medicine ,Cell signaling ,Immunoprecipitation ,Nedd4 Ubiquitin Protein Ligases ,Ubiquitin-Protein Ligases ,Biophysics ,NEDD4 ,macromolecular substances ,immunoprecipitation ,Biochemistry ,testis development ,Mice ,03 medical and health sciences ,Ubiquitin ,Testis ,Animals ,Endosomal Sorting Complexes Required for Transport ,030102 biochemistry & molecular biology ,biology ,Ubiquitination ,Ubiquitin ligase ,Cell biology ,030104 developmental biology ,biology.protein - Abstract
Posttranslational modification by ubiquitination targets proteins for degradation, recycling, stabilization or altered trafficking, and as such can alter cellular signaling pathways. The substrate specificity of this multistep process is controlled by ubiquitin ligases, including those of the HECT domain-containing NEDD4 family. In the testis, ubiquitination of many proteins contributes to organ development and maturation of spermatozoa and NEDD4 is known to be important in the control of spermatogonial stem cell homeostasis. However, a comprehensive understanding of NEDD4 substrates in testis development is lacking. Here we demonstrate high expression of Nedd4 in somatic cells of the mouse testis and in the murine Leydig cell-like cell line TM3. Immunoprecipitation of NEDD4 tagged with GFP at either the amino or carboxyl terminus was subjected to proteomic analysis for interacting proteins. We identified a substantial list of potential interaction partners, including known NEDD4 substrates, proteins involved in ubiquitination and proteins important for testis development and spermatogenesis. We confirmed the interaction of NEDD4 with a subset of these putative interacting proteins, validating the integrity of the dataset. These potential interactors may be further explored to reveal important roles of NEDD4-mediated ubiquitination in the testis. Significance: Ubiquitination is important for testis development and function, and NEDD4 is known to ubiquitinate various proteins to affect cellular signaling and development, including those implicated in spermatogenesis. However, substrates of NEDD4 that are important during testis development remain to be identified. Here we report NEDD4 expression in the developing testis and TM3 testicular cell line. This study identifies a substantial list of NEDD4 interacting proteins in the TM3 testicular cell line, with validation of some of these interactions. Hence, this provides novel NEDD4 targets that may contribute to testis development and function that may be further explored. Refereed/Peer-reviewed
- Published
- 2020
49. The PP2A-Integrator-CDK9 axis fine-tunes transcription and can be targeted therapeutically in cancer.
- Author
-
Vervoort, Stephin J., Welsh, Sarah A., Devlin, Jennifer R., Barbieri, Elisa, Knight, Deborah A., Offley, Sarah, Bjelosevic, Stefan, Costacurta, Matteo, Todorovski, Izabela, Kearney, Conor J., Sandow, Jarrod J., Fan, Zheng, Blyth, Benjamin, McLeod, Victoria, Vissers, Joseph H.A., Pavic, Karolina, Martin, Ben P., Gregory, Gareth, Demosthenous, Elena, and Zethoven, Magnus
- Subjects
- *
RNA polymerase II , *CYCLIN-dependent kinases , *PHOSPHOPROTEIN phosphatases , *TRANSGENIC organisms , *PHOSPHATASES - Abstract
Gene expression by RNA polymerase II (RNAPII) is tightly controlled by cyclin-dependent kinases (CDKs) at discrete checkpoints during the transcription cycle. The pausing checkpoint following transcription initiation is primarily controlled by CDK9. We discovered that CDK9-mediated, RNAPII-driven transcription is functionally opposed by a protein phosphatase 2A (PP2A) complex that is recruited to transcription sites by the Integrator complex subunit INTS6. PP2A dynamically antagonizes phosphorylation of key CDK9 substrates including DSIF and RNAPII-CTD. Loss of INTS6 results in resistance to tumor cell death mediated by CDK9 inhibition, decreased turnover of CDK9 phospho-substrates, and amplification of acute oncogenic transcriptional responses. Pharmacological PP2A activation synergizes with CDK9 inhibition to kill both leukemic and solid tumor cells, providing therapeutic benefit in vivo. These data demonstrate that fine control of gene expression relies on the balance between kinase and phosphatase activity throughout the transcription cycle, a process dysregulated in cancer that can be exploited therapeutically. [Display omitted] • Loss of the INTS6 subunit of Integrator confers resistance to CDK9 inhibition • INTS6 recruits PP2A to chromatin and forms a submodule of Integrator (Int-PP2A) • Int-PP2A opposes CDK9 at the phosphorylation level to fine-tune transcription • PP2A activators synergize therapeutically with CDK9 inhibitors in cancer Interplay between PP2A and CDK9 provides a control point for gene expression that can be exploited to inhibit tumorigenesis. [ABSTRACT FROM AUTHOR]
- Published
- 2021
- Full Text
- View/download PDF
50. High CD123 levels enhance proliferation in response to IL-3, but reduce chemotaxis by downregulating CXCR4 expression
- Author
-
Hayley S. Ramshaw, Gabriela Brumatti, Paul G Ekert, Angel F. Lopez, Jarrod J. Sandow, Melanie K. Pudney, Mara Dottore, Nicole L Wittwer, Richard J D'Andrea, Ceilidh Marchant, Wittwer, Nicole L, Brumatti, Gabriela, Marchant, Ceilidh, Sandow, Jarrod J, Pudney, Melanie K, Dottore, Mara, D'Andrea, Richard J, Lopez, Angel F, Ekert, Paul G, and Ramshaw, Hayley S
- Subjects
0301 basic medicine ,Stromal cell ,Hematopoiesis and Stem Cells ,Myeloid leukemia ,Hematology ,Biology ,leukemic stem cells ,myeloid leukemia ,03 medical and health sciences ,Haematopoiesis ,fetal liver cells ,030104 developmental biology ,Immunophenotyping ,medicine.anatomical_structure ,Immunology ,Cancer research ,medicine ,Bone marrow ,Stem cell ,Signal transduction ,Protein kinase B - Abstract
High expression of the a chain of the interleukin-3 receptor (IL-3R alpha; CD123) is a hallmark of acute myeloid leukemia (AML) leukemic stem cells (LSCs). Elevated CD123 expression is part of the diagnostic immunophenotyping of myeloid leukemia, and higher expression is associated with poor prognosis. However, the biological basis of the poorer prognosis is unclear, and may include heightened IL-3 signaling and non-cell autonomous interactions with the bone marrow (BM) microenvironment. We used TF-1 cells expressing different levels of CD123 and found elevated CD123 levels amplified the proliferative response to exogenous IL-3 and maintained viability in reducing IL-3 concentrations. This was associated with stronger activation of STAT5, Akt, and extracellular signal-regulated kinase 1/2 in vitro. Surprisingly, in vivo e14.5 fetal liver cells transduced with retroviral constructs to express high CD123 failed to engraft in syngeneic recipients. In exploring the underlying mechanism for this, we found that CXCR4, a key molecule involved in LSC/BM interactions, was specifically downregulated in CD123 overexpressing cells in a manner dependent on IL-3 signaling. CXCR4 downregulation was sufficient to alter the chemotactic response of hematopoietic cells to stromal derived factor-1 (SDF-1). Thus, we propose that the overexpression of CD123 in AML LSC dictates their location by altering CXCR4/SDF-1 interaction in the BM, raising the possibility that this mechanism underpins the egress of BM AML LSC and more mature cells into the circulation. Refereed/Peer-reviewed
- Published
- 2017
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