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Ras Guanine Nucleotide Releasing Factor 1 (RasGrf1) Enhancement of Trk Receptor-Mediated Neurite Outgrowth Requires Activation of Both H-Ras and Rac

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Abstract

We previously demonstrated that the guanine nucleotide exchange factor, RasGrf1, binds nerve growth factor (NGF)-activated TrkA and facilitates neurotrophin-induced neurite outgrowth in PC12 cells. RasGrf1 can activate both Ras and Rac, via intrinsic Cdc25 and DH domains, respectively, suggesting that the activation of both could contribute to this process. Previous studies have assayed constitutive neurite outgrowth following RasGrf1 over-expression in PC12 cells, in either the absence or presence of ectopic H-Ras, and have suggested an essential role for either Ras or Rac depending on the presence of H-Ras over-expression. In contrast, in this study, we have addressed the mechanism of how RasGrf1 facilitates neurite outgrowth in response to the neurotrophins, NGF and BDNF. Using Ras/Rac activation assays and site-directed RasGrf1 mutants, we find that both Ras and Rac are essential to neurotrophin-induced neurite outgrowth. Moreover, we find that H-Ras over-expression rescues the loss of neurite outgrowth observed with a Rac minus mutant and that RasGrf1 differentially stimulates NGF-dependent activation of Rac or Ras, depending on cell type. Collectively, these studies clarify the mechanism of how RasGrf1 expression facilitates neurotrophin-induced neurite outgrowth. Moreover, they suggest that H-Ras over-expression should be used with caution to measure phenotypic responses.

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Acknowledgments

This work was supported by a Discovery Grant to Susan O. Meakin from the Natural Sciences and Engineering Research Council of Canada (RGPIN 341947). K. Robinson-Brookes and Asghar Talebian were supported with funds from an Ontario Graduate Scholarship and an Ontario Graduate Scholarship in Science and Technology, respectively.

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Authors and Affiliations

  1. Molecular Brain Research Group, Laboratory of Neural Signaling, Robarts Research Institute, University of Western Ontario, 100 Perth Drive, London, ON, Canada, N6A 5K8

    Asghar Talebian, Kim Robinson-Brookes, James I. S. MacDonald & Susan O. Meakin

  2. Department of Biochemistry, University of Western Ontario, London, ON, Canada, N6A 5C1

    Asghar Talebian & Susan O. Meakin

  3. Graduate Program in Neuroscience, University of Western Ontario, London, ON, Canada, N6A 5C1

    Kim Robinson-Brookes & Susan O. Meakin

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  1. Asghar Talebian
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  2. Kim Robinson-Brookes
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  3. James I. S. MacDonald
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  4. Susan O. Meakin
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Correspondence to Susan O. Meakin.

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Asghar Talebian and Kim Robinson-Brookes are co-first authors.

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Talebian, A., Robinson-Brookes, K., MacDonald, J.I.S. et al. Ras Guanine Nucleotide Releasing Factor 1 (RasGrf1) Enhancement of Trk Receptor-Mediated Neurite Outgrowth Requires Activation of Both H-Ras and Rac. J Mol Neurosci 49, 38–51 (2013). https://doi.org/10.1007/s12031-012-9847-9

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  • DOI: https://doi.org/10.1007/s12031-012-9847-9

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