Back to Search
Start Over
Layilin promotes mitochondrial fission by cyclin-dependent kinase 1 and dynamin-related protein 1 activation in HEK293T cells
- Source :
- Biochemical and biophysical research communications. 549
- Publication Year :
- 2021
-
Abstract
- Object Functions of layilin, a type 1 transmembrane protein with a C-type lectin motif, remain to be clarified. We here investigated precise intracellular localization of layilin and the location-related functions. Methods We used HEK293T cells to assess the co-localization of layilin with different individual organelle markers by double immunostaining. We then investigated mitochondrial morphology in layilin-knockdown (KD) conditions, also with immunostaining. Next, we measured amounts of proteins involved in regulation of mitochondrial dynamics, DRP1, pS616-DRP1, mitofusin1, mitofusin2, CDK1, pY15-CDK1, and cyclin B1, in layilin-KD cells versus control cells by Western blot. Furthermore, by using layilin-knockout (KO) cells, amounts of CDK1 and pY15-CDK1 as well as mitochondrial morphology were investigated. Result We found that layilin localized to mitochondria rather than the other organelles. Small round-shape mitochondria were observed in control cells, whereas elongated and highly connected mitochondria were observed in layilin-KD cells. Amounts of active DRP1 (pS616-DRP1) and total DRP1 were significantly smaller in layilin-KD cells than in controls. Amounts of inactive CDK1 (pY15-CDK1) were significantly larger in layilin-KD cells than in controls. No other tested molecules were significantly altered in layilin-KD cells. Amounts of inactive CDK1 were significantly larger in layilin-KO cells than in wild type (WT) cells. Small round-shape mitochondria were observed in WT cells, whereas elongated and highly connected mitochondria were observed in layilin-KO cells. Conclusion We here demonstrated that layilin played a role in the maintenance of fragmented mitochondria in mitochondrial dynamics and that this function needed CDK1 and DRP1 activation. Our data unveiled a novel function for layilin, regulation of mitochondrial dynamics.
- Subjects :
- 0301 basic medicine
Dynamins
Biophysics
Mitochondrion
Biochemistry
Mitochondrial Dynamics
Models, Biological
03 medical and health sciences
DNM1L
0302 clinical medicine
Organelle
CDC2 Protein Kinase
Humans
Lectins, C-Type
Molecular Biology
Cyclin-dependent kinase 1
Chemistry
HEK 293 cells
Wild type
Cell Biology
Transmembrane protein
Cell biology
Mitochondria
030104 developmental biology
HEK293 Cells
030220 oncology & carcinogenesis
Gene Knockdown Techniques
Mitochondrial fission
Subjects
Details
- ISSN :
- 10902104
- Volume :
- 549
- Database :
- OpenAIRE
- Journal :
- Biochemical and biophysical research communications
- Accession number :
- edsair.doi.dedup.....49196f459b827fe045f8de4dd768ae66