RRC ID 49509
Author Makyio H, Ohgi M, Takei T, Takahashi S, Takatsu H, Katoh Y, Hanai A, Ueda T, Kanaho Y, Xie Y, Shin HW, Kamikubo H, Kataoka M, Kawasaki M, Kato R, Wakatsuki S, Nakayama K.
Title Structural basis for Arf6-MKLP1 complex formation on the Flemming body responsible for cytokinesis.
Journal EMBO J
Abstract A small GTPase, Arf6, is involved in cytokinesis by localizing to the Flemming body (the midbody). However, it remains unknown how Arf6 contributes to cytokinesis. Here, we demonstrate that Arf6 directly interacts with mitotic kinesin-like protein 1 (MKLP1), a Flemming body-localizing protein essential for cytokinesis. The crystal structure of the Arf6-MKLP1 complex reveals that MKLP1 forms a homodimer flanked by two Arf6 molecules, forming a 2:2 heterotetramer containing an extended β-sheet composed of 22 β-strands that spans the entire heterotetramer, suitable for interaction with a concave membrane surface at the cleavage furrow. We show that, during cytokinesis, Arf6 is first accumulated around the cleavage furrow and, prior to abscission, recruited onto the Flemming body via interaction with MKLP1. We also show by structure-based mutagenesis and siRNA-mediated knockdowns that the complex formation is required for completion of cytokinesis. A model based on these results suggests that the Arf6-MKLP1 complex plays a crucial role in cytokinesis by connecting the microtubule bundle and membranes at the cleavage plane.
Volume 31(11)
Pages 2590-603
Published 2012-5-30
DOI 10.1038/emboj.2012.89
PII emboj201289
PMID 22522702
PMC PMC3365427
MeSH ADP-Ribosylation Factor 6 ADP-Ribosylation Factors / chemistry ADP-Ribosylation Factors / genetics ADP-Ribosylation Factors / metabolism* Animals Crystallography, X-Ray Cytokinesis* HeLa Cells Humans Mice Microtubule-Associated Proteins / chemistry Microtubule-Associated Proteins / genetics Microtubule-Associated Proteins / metabolism* Mutation Protein Binding
IF 9.889
Times Cited 35
DNA material pcDNA3/hArf1(WT)-mCherry (RDB15345) pcDNA3/hArf3-mCherry (RDB15346) pcDNA3/hArf5(WT)-mCherry (RDB15347) pEGFP-N3/hArf5(WT)-EGFP (RDB15348) pcDNA3/hArf6(WT)-HA (RDB15349) pcDNA3/hArf6(Q67L)-HA (RDB15350) pcDNA3/hArf6(T27N)-HA (RDB15351) pcDNA3/hArf6(WT)-mCherry (RDB15352) pEGFP-N3/hArf6(WT)-EGFP (RDB15353).