Human mitotic spindle-associated protein PRC1 inhibits MgcRacGAP activity toward Cdc42 during the metaphase

J Biol Chem. 2004 Apr 16;279(16):16394-402. doi: 10.1074/jbc.M313257200. Epub 2004 Jan 26.

Abstract

Although many proteins have been shown to participate in mitotic events, including cytokinesis, their specific roles and interactions remain unclear. A novel interaction of proteins is demonstrated in this report. Yeast two-hybrid screening using PRC1 (protein-regulating cytokinesis 1) cDNA, a human mitotic spindle-associated cyclin-dependent kinase (CDK) substrate, which is involved in cytokinesis, as bait was performed. Data show that the PRC1 bait bound to MgcRacGAP, which is a GTPase-activating protein (GAP) for the Rho family GTPases also involved in cytokinesis. In addition, the two proteins showed similar localization during the M phase. PRC1 was shown to bind to the COOH-terminal GAP-conserved domain of MgcRacGAP and to inhibit its GAP activity toward Cdc42. This binding and/or inhibition of MgcRacGAP GAP activity was found to depend on further binding of PRC1 to the basic region (125-285 amino acids) of MgcRacGAP. Furthermore, the basic region was phosphorylated with Aurora B kinase, and this phosphorylation prevented the inhibition of GAP activity by PRC1. Cells overexpressing a phosphorylation mimic mutant of MgcRacGAP exhibited an abnormality of spindle morphology in the metaphase. Cdc42 showed high activity and was localized to the mitotic spindles and centrosomes during the metaphase. We propose that PRC1 down-regulates the GAP activity of MgcRac-GAP during the metaphase and thereby contributes to the correct formation of the spindle.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Cell Cycle Proteins / metabolism*
  • Down-Regulation
  • GTPase-Activating Proteins / antagonists & inhibitors*
  • HeLa Cells
  • Humans
  • Metaphase / physiology*
  • Protein Binding
  • Signal Transduction
  • Spindle Apparatus / physiology
  • cdc42 GTP-Binding Protein / metabolism*

Substances

  • Cell Cycle Proteins
  • GTPase-Activating Proteins
  • PRC1 protein, human
  • mgcRacGAP
  • cdc42 GTP-Binding Protein