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PMID: 25209408 Published · epublish English Journal Article Research Support, Non-U.S. Gov't

Mechanistic insight into GPCR-mediated activation of the microtubule-associated RhoA exchange factor GEF-H1.

Nature communications ·Vol. 5 ·2014-09-11 ·Pages 4857

Meiri D, Marshall CB, Mokady D, LaRose J, Mullin M, Gingras AC, Ikura M, Rottapel R

Abstract

The RhoGEF GEF-H1 can be sequestered in an inactive state on polymerized microtubules by the dynein motor light-chain Tctex-1. Phosphorylation of GEF-H1 Ser885 by PKA or PAK kinases creates an inhibitory 14-3-3-binding site. Here we show a new mode of GEF-H1 activation in response to the G-protein-coupled receptor (GPCR) ligands lysophosphatidic acid (LPA) or thrombin that is independent of microtubule depolymerization. LPA/thrombin stimulates disassembly of the GEF-H1:dynein multi-protein complex through the concerted action of Gα and Gβγ. Gα binds directly to GEF-H1 and displaces it from Tctex-1, while Gβγ binds to Tctex-1 and disrupts its interaction with the dynein intermediate chain, resulting in the release of GEF-H1. Full activation of GEF-H1 requires dephosphorylation of Ser885 by PP2A, which is induced by thrombin. The coordinated displacement of GEF-H1 from microtubules by G-proteins and its dephosphorylation by PP2A demonstrate a multistep GEF-H1 activation and present a unique mechanism coupling GPCR signalling to Rho activation.

MeSH Terms

14-3-3 Proteins/metabolism Animals Dyneins/metabolism GTP-Binding Protein alpha Subunits, G12-G13/drug effects,metabolism GTP-Binding Protein beta Subunits/drug effects,metabolism GTP-Binding Protein gamma Subunits/drug effects,metabolism Lysophospholipids/pharmacology Mice Mice, Knockout Microtubules/metabolism Phosphorylation Receptors, G-Protein-Coupled/drug effects,metabolism Rho Guanine Nucleotide Exchange Factors/drug effects,metabolism Thrombin/pharmacology p21-Activated Kinases/metabolism

Chemicals

14-3-3 Proteins Arhgef2 protein, mouse Dynlt1b protein, mouse GTP-Binding Protein beta Subunits GTP-Binding Protein gamma Subunits Lysophospholipids Receptors, G-Protein-Coupled Rho Guanine Nucleotide Exchange Factors p21-Activated Kinases Thrombin Dyneins GTP-Binding Protein alpha Subunits, G12-G13 lysophosphatidic acid

Authors & Affiliations

8 authors, click to expand affiliations / ORCID

Meiri David

Department of Biology, Technion Israel Institute of Technology, Haifa 32000, Israel.

Marshall Christopher B

Princess Margaret Cancer Centre, University Health Network, University of Toronto, 101 College Street, Room 12-704, Toronto Medical Discovery Tower, Toronto, Ontario, Canada M5G 1L7.

Mokady Daphna

Princess Margaret Cancer Centre, University Health Network, University of Toronto, 101 College Street, Room 12-704, Toronto Medical Discovery Tower, Toronto, Ontario, Canada M5G 1L7.

LaRose Jose

Princess Margaret Cancer Centre, University Health Network, University of Toronto, 101 College Street, Room 12-704, Toronto Medical Discovery Tower, Toronto, Ontario, Canada M5G 1L7.

Mullin Michael

Lunenfeld-Tanenbaum Research Institute, Mount Sinai Hospital, 600 University Avenue, Room 992A, Toronto, Ontario, Canada M5G 1X5.

Gingras Anne-Claude

1] Lunenfeld-Tanenbaum Research Institute, Mount Sinai Hospital, 600 University Avenue, Room 992A, Toronto, Ontario, Canada M5G 1X5 [2] Department of Molecular Genetics, University of Toronto, Toronto, Ontario, Canada M5S 1A8.

Ikura Mitsuhiko

Rottapel Robert

1] Princess Margaret Cancer Centre, University Health Network, University of Toronto, 101 College Street, Room 12-704, Toronto Medical Discovery Tower, Toronto, Ontario, Canada M5G 1L7 [2] Department of Medical Biophysics, University of Toronto, 1 King's College Circle, Toronto, Ontario, Canada M5S 1A8 [3] Department of Medicine, University of Toronto, 1 King's College Circle, Toronto, Ontario, Canada M5S 1A8 [4] Department of Immunology, University of Toronto, 1 King's College Circle, Toronto, Ontario, Canada M5S 1A8 [5] Division of Rheumatology, St Michael's Hospital, 30 Bond Street, Toronto, Ontario, Canada M5B 1W8.

Article Info

Journal

Nature communications

Abbr.

Nat Commun

ISSN

2041-1723

DOI

10.1038/ncomms5857

Published

2014-09-11

Epub

2014-00-11

Pages

4857

Language

English

Region

England

NLM ID

101528555

Subset

Grants

Canadian Institutes of Health Research · MOP-84314 · Canada

External Links

PubMed source DOI: 10.1038/ncomms5857 View journal details

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Mechanistic insight into GPCR-mediated activation of the microtubule-associated RhoA exchange factor GEF-H1.

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