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Identification of Ser/Thr kinase and Forkhead Associated Domains in Mycobacterium ulcerans: Characterization of Novel Association between Protein Kinase Q and MupFHA
- Source :
- PLoS Neglected Tropical Diseases, PLoS Neglected Tropical Diseases, Public Library of Science, 2014, 8 (11), pp.e3315. ⟨10.1371/journal.pntd.0003315⟩, PLoS Neglected Tropical Diseases, Vol 8, Iss 11, p e3315 (2014)
- Publication Year :
- 2014
- Publisher :
- Public Library of Science (PLoS), 2014.
-
Abstract
- Background Mycobacterium ulcerans, the causative agent of Buruli ulcer in humans, is unique among the members of Mycobacterium genus due to the presence of the virulence determinant megaplasmid pMUM001. This plasmid encodes multiple virulence-associated genes, including mup011, which is an uncharacterized Ser/Thr protein kinase (STPK) PknQ. Methodology/Principal Findings In this study, we have characterized PknQ and explored its interaction with MupFHA (Mup018c), a FHA domain containing protein also encoded by pMUM001. MupFHA was found to interact with PknQ and suppress its autophosphorylation. Subsequent protein-protein docking and molecular dynamic simulation analyses showed that this interaction involves the FHA domain of MupFHA and PknQ activation loop residues Ser170 and Thr174. FHA domains are known to recognize phosphothreonine residues, and therefore, MupFHA may be acting as one of the few unusual FHA-domain having overlapping specificity. Additionally, we elucidated the PknQ-dependent regulation of MupDivIVA (Mup012c), which is a DivIVA domain containing protein encoded by pMUM001. MupDivIVA interacts with MupFHA and this interaction may also involve phospho-threonine/serine residues of MupDivIVA. Conclusions/Significance Together, these results describe novel signaling mechanisms in M. ulcerans and show a three-way regulation of PknQ, MupFHA, and MupDivIVA. FHA domains have been considered to be only pThr specific and our results indicate a novel mechanism of pSer as well as pThr interaction exhibited by MupFHA. These results signify the need of further re-evaluating the FHA domain –pThr/pSer interaction model. MupFHA may serve as the ideal candidate for structural studies on this unique class of modular enzymes.<br />Author Summary Mycobacterium ulcerans is a slow growing pathogen, which is prevalent in many tropical and sub-tropical countries. M. ulcerans possesses unique signaling pathways with only 13 STPK containing genes. This is strikingly different from its closest homolog Mycobacterium marinum and surprisingly closer to the human pathogen, Mycobacterium tuberculosis. PknQ, MupFHA and MupDivIVA are regulatory proteins encoded by the virulence determining plasmid pMUM001 of M. ulcerans. In addition to characterizing the STPK, we focused on deciphering the basis of interaction between the three partner proteins leading to the identification of critical residues. Present study describes the newly identified phosphoserine-based interactions, which is unique amongst the FHA-domain containing proteins. We confirmed our results using structural analysis via specific mutants and their interaction profiles. Importantly, these data highlight the significance of FHA domains and their role in understanding cellular signaling. This work will encourage further studies to elucidate role of M. ulcerans signaling systems. It will also raise questions like how less studied tropical bacterial pathogens acquire eukaryotic-like Ser/Thr protein kinase and exhibit unusual mechanisms to interact with its partner domains.
- Subjects :
- Proteomics
[SDV]Life Sciences [q-bio]
Protein domains
Plasma protein binding
Biochemistry
Serine
Protein structure
Sequence alignment
Medicine and Health Sciences
Phosphorylation
In vitro kinase assay
Enzyme Chemistry
0303 health sciences
lcsh:Public aspects of medicine
Autophosphorylation
Forkhead Transcription Factors
Recombinant Proteins
Enzymes
Bacterial Pathogens
Cell biology
[SDV.MP]Life Sciences [q-bio]/Microbiology and Parasitology
Infectious Diseases
Medical Microbiology
Protein Binding
Research Article
Protein Structure
lcsh:Arctic medicine. Tropical medicine
lcsh:RC955-962
Protein domain
Molecular Dynamics Simulation
Protein Serine-Threonine Kinases
Biology
Microbiology
Protein–protein interaction
Enzyme Regulation
03 medical and health sciences
Bacterial Proteins
Protein Interactions
Protein kinase A
Microbial Pathogens
030304 developmental biology
Mycobacterium ulcerans
030306 microbiology
Public Health, Environmental and Occupational Health
Computational Biology
Biology and Life Sciences
Proteins
lcsh:RA1-1270
Mycobacterium tuberculosis
[SDV.MP.BAC]Life Sciences [q-bio]/Microbiology and Parasitology/Bacteriology
Protein Structure, Tertiary
Regulatory Proteins
Emerging Infectious Diseases
Enzyme Structure
Enzymology
Subjects
Details
- ISSN :
- 19352735 and 19352727
- Volume :
- 8
- Database :
- OpenAIRE
- Journal :
- PLoS Neglected Tropical Diseases
- Accession number :
- edsair.doi.dedup.....cd618a9258bcf0560dd3da7e2efd4344
- Full Text :
- https://doi.org/10.1371/journal.pntd.0003315