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Anillin Phosphorylation Controls Timely Membrane Association and Successful Cytokinesis
- Source :
- PLoS Genetics, PLoS Genetics, Vol 13, Iss 1, p e1006511 (2017)
- Publication Year :
- 2017
- Publisher :
- Public Library of Science, 2017.
-
Abstract
- During cytokinesis, a contractile ring generates the constricting force to divide a cell into two daughters. This ring is composed of filamentous actin and the motor protein myosin, along with additional structural and regulatory proteins, including anillin. Anillin is a required scaffold protein that links the actomyosin ring to membrane and its organizer, RhoA. However, the molecular basis for timely action of anillin at cytokinesis remains obscure. Here, we find that phosphorylation regulates efficient recruitment of human anillin to the equatorial membrane. Anillin is highly phosphorylated in mitosis, and is a substrate for mitotic kinases. We surveyed function of 46 residues on anillin previously found to be phosphorylated in human cells to identify those required for cytokinesis. Among these sites, we identified S635 as a key site mediating cytokinesis. Preventing S635 phosphorylation adjacent to the AH domain disrupts anillin concentration at the equatorial cortex at anaphase, whereas a phosphomimetic mutant, S635D, partially restores this localization. Time-lapse videomicroscopy reveals impaired recruitment of S635A anillin to equatorial membrane and a transient unstable furrow followed by ultimate failure in cytokinesis. A phosphospecific antibody confirms phosphorylation at S635 in late cytokinesis, although it does not detect phosphorylation in early cytokinesis, possibly due to adjacent Y634 phosphorylation. Together, these findings reveal that anillin recruitment to the equatorial cortex at anaphase onset is enhanced by phosphorylation and promotes successful cytokinesis.<br />Author Summary Human diseases such as cancer and congenital trisomies arise from loss of genetic material during cell division. Yet in most divisions, cells preserve their genetic integrity by strict coordination of cell membrane cleavage (cytokinesis), with accurate separation of genetic material (mitosis). Thus, understanding how mitosis and cytokinesis are coordinated can provide insight into human disease. Anillin is one central integrator of cytokinesis that is recruited in a ring-like structure on the membrane of dividing cells. Protein phosphorylation is a common mechanism regulating timing of events in mitosis; although 46 phosphorylation sites have been mapped on anillin, their functional significance is unknown. Here, we evaluated the effect of blocking anillin phosphorylation on human cell division. Surprisingly, most phosphorylation events appeared dispensable for cytokinesis in the assay used. By contrast, phosphorylation of S635 is important for early recruitment of anillin to the midzone membrane, furrow stabilization and efficient cytokinesis. Our findings highlight a central mechanism regulating the timing of human cytokinesis.
- Subjects :
- 0301 basic medicine
Scaffold protein
Cancer Research
RHOA
Physiology
Cell Membranes
Cultured tumor cells
Biochemistry
0302 clinical medicine
Immune Physiology
Medicine and Health Sciences
Cell Cycle and Cell Division
Post-Translational Modification
Phosphorylation
Genetics (clinical)
Anaphase
Immune System Proteins
Chromosome Biology
Microfilament Proteins
Cell biology
Precipitation Techniques
Cell Processes
Cell lines
Cellular Structures and Organelles
Biological cultures
Research Article
lcsh:QH426-470
Immunology
Mitosis
macromolecular substances
Biology
Research and Analysis Methods
Filamentous actin
Antibodies
Motor protein
03 medical and health sciences
Genetics
Immunoprecipitation
Humans
HeLa cells
Molecular Biology
Ecology, Evolution, Behavior and Systematics
Cytokinesis
Cell Membrane
Biology and Life Sciences
Proteins
Cell Biology
Cell cultures
lcsh:Genetics
030104 developmental biology
Mutation
biology.protein
Protein Processing, Post-Translational
030217 neurology & neurosurgery
Subjects
Details
- Language :
- English
- ISSN :
- 15537404 and 15537390
- Volume :
- 13
- Issue :
- 1
- Database :
- OpenAIRE
- Journal :
- PLoS Genetics
- Accession number :
- edsair.doi.dedup.....5fb8dd27282e42b72d5cbbfb7a9b2daa