Structural determinants of RGS-RhoGEF signaling critical to Entamoeba histolytica pathogenesis

Dustin E Bosch; Adam J Kimple; Alyssa J Manning; Robin E Muller; Francis S Willard; Mischa Machius; Stephen L Rogers; David P Siderovski; Argonne National Lab. (ANL), Argonne, IL (United States). Advanced Photon Source (APS)

doi:10.1016/j.str.2012.11.012

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Structural determinants of RGS-RhoGEF signaling critical to Entamoeba histolytica pathogenesis

Journal article

Open access

Peer reviewed

Structural determinants of RGS-RhoGEF signaling critical to Entamoeba histolytica pathogenesis

Dustin E Bosch, Adam J Kimple, Alyssa J Manning, Robin E Muller, Francis S Willard, Mischa Machius, Stephen L Rogers, David P Siderovski and Argonne National Lab. (ANL), Argonne, IL (United States). Advanced Photon Source (APS)

Structure (London), Vol.21(1), pp.65-75

01/08/2013

DOI: 10.1016/j.str.2012.11.012

PMCID: PMC3545058

PMID: 23260656

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Abstract

G protein signaling pathways, as key components of physiologic responsiveness and timing, are frequent targets for pharmacologic intervention. Here, we identify an effector for heterotrimeric G protein α subunit (EhGα1) signaling from Entamoeba histolytica, the causative agent of amoebic colitis. EhGα1 interacts with this effector and guanosine triphosphatase-accelerating protein, EhRGS-RhoGEF, in a nucleotide state-selective fashion. Coexpression of EhRGS-RhoGEF with constitutively active EhGα1 and EhRacC leads to Rac-dependent spreading in Drosophila S2 cells. EhRGS-RhoGEF overexpression in E. histolytica trophozoites leads to reduced migration toward serum and lower cysteine protease activity, as well as reduced attachment to, and killing of, host cells. A 2.3 Å crystal structure of the full-length EhRGS-RhoGEF reveals a putative inhibitory helix engaging the Dbl homology domain Rho-binding surface and the pleckstrin homology domain. Mutational analysis of the EhGα1/EhRGS-RhoGEF interface confirms a canonical "regulator of G protein signaling" domain rather than a RhoGEF-RGS ("rgRGS") domain, suggesting a convergent evolution toward heterotrimeric and small G protein cross-talk.

Amino Acid Substitution

Animals

Binding Sites

Cell Adhesion

Cell Line

Cell Shape

Cell Survival

Chemotaxis

Crystallography, X-Ray

Drosophila melanogaster

Entamoeba histolytica - metabolism

Entamoeba histolytica - physiology

GTP-Binding Protein alpha Subunits - chemistry

Guanine Nucleotide Exchange Factors - chemistry

Guanine Nucleotide Exchange Factors - genetics

Guanine Nucleotide Exchange Factors - metabolism

Guanosine Triphosphate - chemistry

Host-Parasite Interactions

Hydrolysis

Models, Molecular

Mutagenesis, Site-Directed

Protein Binding

Protein Interaction Domains and Motifs

Protein Structure, Secondary

Protozoan Proteins - chemistry

Protozoan Proteins - genetics

Protozoan Proteins - metabolism

Rho Guanine Nucleotide Exchange Factors

Signal Transduction

Trophozoites - metabolism

Trophozoites - physiology

Details

Title: Subtitle: Structural determinants of RGS-RhoGEF signaling critical to Entamoeba histolytica pathogenesis
Creators: Dustin E Bosch - University of North Carolina at Chapel Hill
Adam J Kimple - University of North Carolina at Chapel Hill
Alyssa J Manning - University of North Carolina at Chapel Hill
Robin E Muller - University of North Carolina at Chapel Hill
Francis S Willard - University of North Carolina at Chapel Hill
Mischa Machius - University of North Carolina at Chapel Hill
Stephen L Rogers - University of North Carolina at Chapel Hill
David P Siderovski - West Virginia University
Argonne National Lab. (ANL), Argonne, IL (United States). Advanced Photon Source (APS)
Resource Type: Journal article
Publication Details: Structure (London), Vol.21(1), pp.65-75
DOI: 10.1016/j.str.2012.11.012
PMID: 23260656
PMCID: PMC3545058
ISSN: 0969-2126
eISSN: 1878-4186
Grant note: R01 GM082892 / NIGMS NIH HHS MH074266 / NIMH NIH HHS GM082892 / NIGMS NIH HHS F30 DK091978 / NIDDK NIH HHS F30 MH074266 / NIMH NIH HHS DK091978 / NIDDK NIH HHS T32 GM008719 / NIGMS NIH HHS
Language: English
Date published: 01/08/2013
Academic Unit: Pathology
Record Identifier: 9984199920802771

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