A conserved, N-terminal tyrosine signal directs Ras for inhibition by Rabex-5

Autoři: Chalita Washington aff001;  Rachel Chernet aff001;  Rewatee H. Gokhale aff001;  Yesenia Martino-Cortez aff001;  Hsiu-Yu Liu aff006;  Ashley M. Rosenberg aff001;  Sivan Shahar aff001;  Cathie M. Pfleger aff001
Působiště autorů: Department of Oncological Sciences, The Icahn School of Medicine at Mount Sinai, New York, New York, United States of America aff001;  University of Cincinnati College of Medicine, Cincinnati, Ohio, United States of America aff002;  The Graduate School of Biomedical Sciences, The Icahn School of Medicine at Mount Sinai, New York, New York, United States of America aff003;  The Tisch Cancer Institute, The Icahn School of Medicine at Mount Sinai, New York, New York, United States of America aff004;  Tufts University School of Medicine, Boston, Massachusetts, United States of America aff005;  Memorial Sloan Kettering Cancer Center, New York, New York, United States of America aff006;  Columbia University, New York, New York, United States of America aff007;  New York Medical College, Valhalla, New York, United States of America aff008
Vyšlo v časopise: A conserved, N-terminal tyrosine signal directs Ras for inhibition by Rabex-5. PLoS Genet 16(6): e32767. doi:10.1371/journal.pgen.1008715
Kategorie: Research Article
doi: https://doi.org/10.1371/journal.pgen.1008715


Dysregulation of the Ras oncogene in development causes developmental disorders, “Rasopathies,” whereas mutational activation or amplification of Ras in differentiated tissues causes cancer. Rabex-5 (also called RabGEF1) inhibits Ras by promoting Ras mono- and di-ubiquitination. We report here that Rabex-5-mediated Ras ubiquitination requires Ras Tyrosine 4 (Y4), a site of known phosphorylation. Ras substitution mutants insensitive to Y4 phosphorylation did not undergo Rabex-5-mediated ubiquitination in cells and exhibited Ras gain-of-function phenotypes in vivo. Ras Y4 phosphomimic substitution increased Rabex-5-mediated ubiquitination in cells. Y4 phosphomimic substitution in oncogenic Ras blocked the morphological phenotypes associated with oncogenic Ras in vivo dependent on the presence of Rabex-5. We developed polyclonal antibodies raised against an N-terminal Ras peptide phosphorylated at Y4. These anti-phospho-Y4 antibodies showed dramatic recognition of recombinant wild-type Ras and RasG12V proteins when incubated with JAK2 or SRC kinases but not of RasY4F or RasY4F,G12V recombinant proteins suggesting that JAK2 and SRC could promote phosphorylation of Ras proteins at Y4 in vitro. Anti-phospho-Y4 antibodies also showed recognition of RasG12V protein, but not wild-type Ras, when incubated with EGFR. A role for JAK2, SRC, and EGFR (kinases with well-known roles to activate signaling through Ras), to promote Ras Y4 phosphorylation could represent a feedback mechanism to limit Ras activation and thus establish Ras homeostasis. Notably, rare variants of Ras at Y4 have been found in cerebellar glioblastomas. Therefore, our work identifies a physiologically relevant Ras ubiquitination signal and highlights a requirement for Y4 for Ras inhibition by Rabex-5 to maintain Ras pathway homeostasis and to prevent tissue transformation.

Klíčová slova:

Carcinogenesis – Drosophila melanogaster – Eyes – Phenotypes – Phosphorylation – Ras signaling – RNA interference – Ubiquitination


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