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Experimental Cell Research
Volume 293, Issue 2, 15 February 2004, Pages 275-282
 
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doi:10.1016/j.yexcr.2003.10.015    How to Cite or Link Using DOI (Opens New Window)
Copyright © 2003 Elsevier Inc. All rights reserved.

MgcRacGAP regulates cortical activity through RhoA during cytokinesis*1

Jae-Seon Leea, Corresponding Author Contact Information, E-mail The Corresponding Author, Keiju Kamijoa, Naoya Oharaa, Toshio Kitamurab and Toru Mikia

a Molecular Tumor Biology Section, Basic Research Laboratory, National Cancer Institute, Bethesda, MD 20892-4255, USA b Division of Hematopoietic Factors, The Institute of Medical Science, University of Tokyo, Minato, Tokyo 108-8639, Japan

Received 25 May 2003; 
revised 9 September 2003. 
Available online 27 November 2003.

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Abstract

Although Rho GTPases regulate multiple cellular events, their role in cell division is still obscure. Here we show that expression of a GTPase-activating protein (GAP)-deficient mutant (R386A) of the Rho regulator MgcRacGAP induces abnormal cortical activity during cytokinesis in U2OS cells. Multiple large blebs were observed in cells expressing MgcRacGAP R386A from the onset of anaphase to the late stage of cell division. When mitotic blebbing was excessive, cytokinesis was inhibited, and cells with micronuclei were generated. It has been reported that blebbing is caused by abnormal cortical activity. The MgcRacGAP R386A-induced abnormal cortical activity was inhibited by the dominant negative form of RhoA, but not Rac1 or Cdc42. Moreover, expression of constitutively active RhoA also induced drastic cortical activity during cytokinesis. Unlike apoptotic blebbing, MgcRacGAP R386A-induced blebbing was not inhibited by the ROCK inhibitor Y-27632, suggesting that MgcRacGAP regulates cortical activity during cytokinesis through a novel signaling pathway. We propose that MgcRacGAP plays a pivotal role in cytokinesis by regulating cortical movement through RhoA.

Author Keywords: MgcRacGAP; RhoA; Cytokinesis; Cortical activity

Article Outline

• Introduction
• Materials and methods
• DNA constructs
• Cell culture and transfection
• Time-lapse video microscopy
• Treatment of Y-27632
• Results
• GAP-deficient MgcRacGAP induces multiple blebs during cytokinesis
• DN-RhoA inhibits MgcRacGAP R386A-induced abnormal cortical activity
• Constitutively active RhoA induces abnormal cortical activity during cytokinesis
• ROCK inhibitor Y27632 does not inhibit MgcRacGAP R386A-induced abnormal cortical activity
• Discussion
• Acknowledgements
• References









Experimental Cell Research
Volume 293, Issue 2, 15 February 2004, Pages 275-282
 
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