A complex of p190RhoGAP-A and anillin modulates RhoA-GTP and the cytokinetic furrow in human cells

Arkadi Manukyan; Kirsten Ludwig; Sergio Sanchez-Manchinelly; Sarah J Parsons; P Todd Stukenberg

doi:10.1242/jcs.151647

A complex of p190RhoGAP-A and anillin modulates RhoA-GTP and the cytokinetic furrow in human cells

J Cell Sci. 2015 Jan 1;128(1):50-60. doi: 10.1242/jcs.151647. Epub 2014 Oct 29.

Authors

Arkadi Manukyan¹, Kirsten Ludwig², Sergio Sanchez-Manchinelly³, Sarah J Parsons⁴, P Todd Stukenberg⁵

Affiliations

¹ Department of Biochemistry and Molecular Genetics, University of Virginia School of Medicine, PO Box 800733, Charlottesville, VA 22908, USA Cancer Center, University of Virginia School of Medicine, PO Box 800733, Charlottesville, VA 22908, USA.
² Cancer Center, University of Virginia School of Medicine, PO Box 800733, Charlottesville, VA 22908, USA Department of Psychiatry and Behavioral Sciences and Jonsson Cancer Center, UCLA, Los Angeles, CA 90095-6900, USA Department of Microbiology, Immunology, and Cancer Biology, University of Virginia School of Medicine, PO Box 800733, Charlottesville, VA 22908, USA.
³ Cancer Center, University of Virginia School of Medicine, PO Box 800733, Charlottesville, VA 22908, USA Department of Microbiology, Immunology, and Cancer Biology, University of Virginia School of Medicine, PO Box 800733, Charlottesville, VA 22908, USA Beckman Corporation, Los Angeles, CA 90025, USA.
⁴ Cancer Center, University of Virginia School of Medicine, PO Box 800733, Charlottesville, VA 22908, USA Department of Microbiology, Immunology, and Cancer Biology, University of Virginia School of Medicine, PO Box 800733, Charlottesville, VA 22908, USA.
⁵ Department of Biochemistry and Molecular Genetics, University of Virginia School of Medicine, PO Box 800733, Charlottesville, VA 22908, USA Cancer Center, University of Virginia School of Medicine, PO Box 800733, Charlottesville, VA 22908, USA pts7h@virginia.edu.

Abstract

The cytokinetic furrow is organized by the RhoA GTPase, which recruits actin and myosin II to the furrow and drives contractility. Here, we show that the RhoA GTPase-activting protein (GAP) p190RhoGAP-A (also known as ARHGAP35) has a role in cytokinesis and is involved in regulating levels of RhoA-GTP and contractility. Cells depleted of p190RhoGAP-A accumulate high levels of RhoA-GTP and markers of high RhoA activity in the furrow, resulting in failure of the cytokinetic furrow to progress to abscission. The loss of p190RhoGAP-A can be rescued by a low dose of the myosin II inhibitor blebbistatin, suggesting that cells fail cytokinesis because they have too much myosin activity. p190RhoGAP-A binds the cytokinetic organizer anillin, and mutants of p190RhoGAP-A that are unable to bind anillin or unable to inactivate RhoA fail to rescue cytokinesis defects in p190RhoGAP-A-depleted cells. Taken together, these data demonstrate that a complex of p190RhoGAP-A and anillin modulates RhoA-GTP levels in the cytokinetic furrow to ensure progression of cytokinesis.

Keywords: Anillin; Cytokinesis; Myosin II; Rho; p190RhoGAP.

Publication types

Research Support, N.I.H., Extramural
Research Support, Non-U.S. Gov't

MeSH terms

Cytokinesis / drug effects
Cytokinesis / physiology*
Guanine Nucleotide Exchange Factors / genetics
Guanine Nucleotide Exchange Factors / metabolism*
HeLa Cells
Heterocyclic Compounds, 4 or More Rings
Humans
Microfilament Proteins / genetics
Microfilament Proteins / metabolism*
Mutation
Protein Binding / drug effects
Protein Binding / physiology
Repressor Proteins / genetics
Repressor Proteins / metabolism*
rhoA GTP-Binding Protein / genetics
rhoA GTP-Binding Protein / metabolism*

Substances

ANLN protein, human
ARHGAP35 protein, human
Guanine Nucleotide Exchange Factors
Heterocyclic Compounds, 4 or More Rings
Microfilament Proteins
Repressor Proteins
RHOA protein, human
blebbistatin
rhoA GTP-Binding Protein

Abstract

Publication types

MeSH terms

Substances

Grants and funding