Molecular and functional characterization of a Rho GDP dissociation inhibitor in the filamentous fungus Tuber borchii.

BMC Microbiology
M MenottaF Rivero

Abstract

Small GTPases of the Rho family function as tightly regulated molecular switches that govern important cellular functions in eukaryotes. Several families of regulatory proteins control their activation cycle and subcellular localization. Members of the guanine nucleotide dissociation inhibitor (GDI) family sequester Rho GTPases from the plasma membrane and keep them in an inactive form. We report on the characterization the RhoGDI homolog of Tuber borchii Vittad., an ascomycetous ectomycorrhizal fungus. The Tbgdi gene is present in two copies in the T. borchii genome. The predicted amino acid sequence shows high similarity to other known RhoGDIs. Real time PCR analyses revealed an increased expression of Tbgdi during the phase preparative to the symbiosis instauration, in particular after stimulation with root exudates extracts, that correlates with expression of Tbcdc42. In a translocation assay TbRhoGDI was able to solubilize TbCdc42 from membranes. Surprisingly, TbRhoGDI appeared not to interact with S. cerevisiae Cdc42, precluding the use of yeast as a surrogate model for functional studies. To study the role of TbRhoGDI we performed complementation experiments using a RhoGDI null strain of Dictyostelium discoideum, a model...Continue Reading

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Citations

Apr 15, 2009·Antimicrobial Agents and Chemotherapy·Mark E ShirtliffMary Ann Jabra-Rizk
Sep 27, 2012·Canadian Journal of Microbiology·Lucia PotenzaVilberto Stocchi
Dec 13, 2012·International Journal of Oncology·Yong-Sheng JiangTakahide Ota
May 24, 2012·Biotechnology & Genetic Engineering Reviews·Marjatta RaudaskoskiJ Stephen Horton
Dec 1, 2017·Applied Microbiology and Biotechnology·Giada Giusi PicceriLaura Chiarantini

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Datasets Mentioned

BETA
EU044761
AAF70843
AAC17610
AAF21198

Methods Mentioned

BETA
GTPases
nucleotide exchange
two-hybrid
GTPase
PCR
GTPses
transfection
two hybrid

Software Mentioned

ClustalX
SeqPup
SUN JAVA VM
BLAST

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