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Early endosome localization and activity of RasGEF1b, a toll-like receptor-inducible Ras guanine-nucleotide exchange factor

Genes & Immunity volume 11, pages 447–457 (2010)Cite this article

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Abstract

Guanine-nucleotide exchange factors (GEFs) stimulate the intrinsic GDP/GTP exchange activity of Ras and promote the formation of active Ras–GTP, which in turn controls diverse signalling networks important for the regulation of cell proliferation, survival, differentiation, vesicular trafficking, and gene expression. RasGEF1b is a GEF, whose expression is induced in macrophages on stimulation with toll-like receptor (TLR) agonists. Here, we showed that in vitro RasGEF1b expression by macrophages is mostly induced by TLR3 (poly I:C) and TLR4 (lipopolysaccharyde) through the MyD88-independent pathway. In vivo infection with the protozoan parasites Trypanosoma cruzi and Plasmodium chabaudi induced RasGEF1b in an MyD88-, TRIF-, and IFN-γ-dependent manner. Ectopically expressed RasGEF1b was found, mostly, in the heavy membrane fraction of HEK 293T, and by confocal microscopy, it was found to be located at early endosomes. Computational modelling of the RasGEF1b–Ras interaction revealed that RasGEF1b interacts with the binding domain site of Ras, a critical region for interacting with GEFs involved in the activation of Ras-Raf-MEK-ERK pathway. More important, RasGEF1b was found to be closely associated with Ras in live cells and to trigger Ras activity. Altogether, these results indicate that on TLR activation, RasGEF1b may trigger Ras-like proteins and regulate specific biological activities described for this subtype of GTPases.

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Acknowledgements

We are grateful to Ronaldo Nagem for help and advice regarding the virtual analysis of RasGEF1b. This study was supported by FAPEMIG, the National Institute of Science and Technology for Vaccine Development/CNPq and the Ludwig Institute of Cancer Research—Atlantic Philanthropy—Program for Clinical Discovery. RTG and AMS are research fellows of CNPq.

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Author notes

  1. T N Silveira

    Present address: 6Current address: Department of Cell Biology, School of Medicine from Ribeirão Preto, University of São Paulo, Av. Bandeirantes 3900, Ribeirão Preto, São Paulo, Brazil.,

  2. W A Andrade and A M Silva: These authors contributed equally to this work.

Authors and Affiliations

  1. Department of Biochemistry and Immunology, Federal University of Minas Gerais, Minas Gerais, Brazil

    W A Andrade, A P C Salgado, H M Andrade & R T Gazzinelli

  2. Department of Morphology, Federal University of Minas Gerais, Minas Gerais, Brazil

    A M Silva & F V Dall'Orto

  3. René Rachou Research Center, Oswaldo Cruz Foundation, Belo Horizonte, Minas Gerais, Brazil

    A M Silva, V Souza Alves, S A Garcia, T N Silveira & R T Gazzinelli

  4. University of Massachusetts Medical School, Worcester, MA, USA

    M B Melo & R T Gazzinelli

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  1. W A Andrade
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Andrade, W., Silva, A., Alves, V. et al. Early endosome localization and activity of RasGEF1b, a toll-like receptor-inducible Ras guanine-nucleotide exchange factor. Genes Immun 11, 447–457 (2010). https://doi.org/10.1038/gene.2009.107

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