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

Current biology : CB (2020-03-21)
Xi Chen, Kangji Wang, Tatyana Svitkina, Erfei Bi
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.

MATERIALS
Product Number
Brand
Product Description

Sigma-Aldrich
L-Methionine, reagent grade, ≥98% (HPLC)
Sigma-Aldrich
L-Threonine, reagent grade, ≥98% (HPLC)
Sigma-Aldrich
Sodium chloride, puriss. p.a., ACS reagent, reag. ISO, reag. Ph. Eur., ≥99.5%
Millipore
ReBlot Plus Kit