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Critical Roles of a RhoGEF-Anillin Module in Septin Architectural Remodeling during Cytokinesis.

ABSTRACT: How septin architecture is remodeled from an hourglass to a double ring during cytokinesis in fungal and animal cells remains unknown. Here, we show that during the hourglass-to-double-ring transition in budding yeast, septins acquire a "zonal architecture" in which paired septin filaments that are organized along the mother-bud axis associate with circumferential single septin filaments, the Rho guanine-nucleotide-exchange factor (RhoGEF) Bud3, and the anillin-like protein Bud4 exclusively at the outer zones and with myosin-II filaments in the middle zone. Deletion of Bud3 or its Bud4-interacting domain, but not its RhoGEF domain, leads to a complete loss of the single filaments, whereas deletion of Bud4 or its Bud3-interacting domain destabilizes the transitional hourglass, especially at the mother side, with partial loss of both filament types. Deletion of Bud3 and Bud4 together further weakens the transitional structure and abolishes the double ring formation while causing no obvious defect in actomyosin ring constriction. This and further analyses suggest that Bud3 stabilizes the single filaments, whereas Bud4 strengthens the interaction between the paired and single filaments at the outer zones of the transitional hourglass, as well as in the double ring. This study reveals a striking zonal architecture for the transitional hourglass that pre-patterns two cytokinetic structures-a septin double ring and an actomyosin ring-and also defines the essential roles of a RhoGEF-anillin module in septin architectural remodeling during cytokinesis at the filament level.

SUBMITTER: Chen X

PROVIDER: S-EPMC7176533 | biostudies-literature |

REPOSITORIES: biostudies-literature

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An anillin homologue, Mid2p, acts during fission yeast cytokinesis to organize the septin ring and promote cell separation.

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| S-EPMC3325936 | biostudies-literature

Anillin Phosphorylation Controls Timely Membrane Association and Successful Cytokinesis.

Project description: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.

| S-EPMC5230765 | biostudies-literature

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