Interaction between Anillin and RacGAP50C connects the actomyosin contractile ring with spindle microtubules at the cell division site.
Anillin, one of the first factors recruited to the cleavage site during cytokinesis, interacts with actin, myosin II and septins, and is essential for proper organization of the actomyosin contractile ring. We employed affinity-purification methodology coupled with mass spectrometry to identify Anillin-interacting molecules in Drosophila cells. We isolated several actin and myosin proteins, three of the five Drosophila septins and RacGAP50C (Tum), a component of the centralspindlin complex. Using drug and RNA interference (RNAi) treatments we established that F-actin is essential for Anillin cortical localization in prometaphase but not for its accumulation at the cleavage furrow after anaphase onset. Moreover, septins were not recruited to the cleavage site in cells in which Anillin was knocked down by RNAi, but localized to central-spindle microtubules, suggesting that septins travel along microtubules to interact with Anillin at the furrow. Finally, we demonstrate that RacGAP50C is necessary for Anillin accumulation at the furrow and that the two proteins colocalize in vivo and interact in vitro. Thus, in addition to its role in activating RhoA signalling, RacGAP50C also controls the proper assembly of the actomyosin ring by interacting with Anillin at the cleavage furrow.
D'Avino
Pier Paolo
PP
Cancer Research UK Cell Cycle Genetics Research Group, Department of Genetics, University of Cambridge, Downing Street, Cambridge, CB2 3EH, UK. p.davino@gen.cam.ac.uk
Takeda
Tetsuya
T
Capalbo
Luisa
L
Zhang
Wei
W
Lilley
Kathryn S
KS
Laue
Ernest D
ED
Glover
David M
DM
D000205:Actomyosin / Q000378:metabolism*; D000818:Animals; D002455:Cell Division; D002460:Cell Line; D003285:Contractile Proteins / Q000378:metabolism*; D004330:Drosophila; D029721:Drosophila Proteins / Q000378:metabolism*; D020690:GTPase-Activating Proteins / Q000378:metabolism*; D007150:Immunohistochemistry; D008870:Microtubules / Q000378:metabolism*; D011485:Protein Binding
D003285:Contractile Proteins; D029721:Drosophila Proteins; D020690:GTPase-Activating Proteins; C095886:anillin; C502951:tum protein, Drosophila; D000205:Actomyosin