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Treatment of Eker rats (<jats:italic>Tsc\u20102<\/jats:italic><jats:sup>EK\/+<\/jats:sup>), bearing a mutation in one allele of the tuberous sclerosis\u20102 (<jats:italic>Tsc\u20102<\/jats:italic>) gene, with the nephrocarcinogen 2,3,5\u2010<jats:italic>tris<\/jats:italic>\u2010(glutathion\u2010S\u2010yl) hydroquinone (TGHQ) results in loss of the wild\u2010type allele of <jats:italic>Tsc\u20102<\/jats:italic> in renal preneoplastic lesions and tumors. These tumors have increased protein expression of B\u2010Raf, C\u2010Raf (Raf\u20101), and increased expression and activity of ERK kinase. Similar changes are observed in Raf kinases following TGHQ\u2010mediated transformation of primary renal epithelial cells derived from <jats:italic>Tsc\u20102<\/jats:italic><jats:sup>EK\/+<\/jats:sup> rats (QTRRE cells), cells that are also null for tuberin. Herein, we utilized LC\u2010MS\/MS to identify constitutive phosphorylation of S345 and S483 in both 100\u2010 and 95\u2010kDa forms of B\u2010Raf in QTRRE cells. Using microRotofor liquid\u2010phase isoelectric focusing, we identified four fractions of B\u2010Raf that contain different post\u2010translational modification profiles in QTRRE cells. Amplification of the kinase domain of B\u2010Raf from QTRRE cells, outer\u2010stripe of the outer medulla of 8\u2010month TGHQ\u2010 or vehicle\u2010treated <jats:italic>Tsc\u20102<jats:sup>+\/+<\/jats:sup><\/jats:italic>and <jats:italic>Tsc\u20102<\/jats:italic><jats:sup>EK\/+<\/jats:sup> rats, as well as tumors excised from 8\u2010month TGHQ\u2010treated <jats:italic>Tsc\u20102<\/jats:italic><jats:sup>EK\/+<\/jats:sup> rats revealed three splice variants of B\u2010Raf within the kinase domain. These splice variants differed by approximately 340, 544, and 600 bp; confirmed by sequencing. No point mutations within the kinase domain of B\u2010Raf were identified. In addition, B\u2010Raf\/Raf\u20101\/14\u20103\u20103 complex formation in the QTRRE cells was decreased by sorafenib, with concomitant selective decreases in p\u2010ERK levels. Transcriptional and post\u2010translational characterization of critical kinases, such as B\u2010Raf, may contribute to the progression of tuberous sclerosis RCC. (246\/250) \u00a9 2015 Wiley Periodicals, Inc.<\/jats:p><\/jats:sec>","DOI":"10.1002\/mc.22366","type":"journal-article","created":{"date-parts":[[2015,9,2]],"date-time":"2015-09-02T17:39:10Z","timestamp":1441215550000},"page":"1243-1250","source":"Crossref","is-referenced-by-count":0,"title":["Transcriptional and post\u2010translational modifications of B\u2010Raf in quinol\u2010thioether induced tuberous sclerosis renal cell carcinoma"],"prefix":"10.1002","volume":"55","author":[{"given":"Jennifer D.","family":"Cohen","sequence":"first","affiliation":[{"name":"Southwest Environmental Health Sciences Center College of Pharmacy, The University of Arizona Tucson Arizona"},{"name":"Department of Pharmacology and Toxicology College of Pharmacy, The University of Arizona Tucson Arizona"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Matthew","family":"Labenski","sequence":"additional","affiliation":[{"name":"Southwest Environmental Health Sciences Center College of Pharmacy, The University of Arizona Tucson Arizona"},{"name":"Department of Pharmacology and Toxicology College of Pharmacy, The University of Arizona Tucson Arizona"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Nicholas J.","family":"Mastrandrea","sequence":"additional","affiliation":[{"name":"Southwest Environmental Health Sciences Center College of Pharmacy, The University of Arizona Tucson Arizona"},{"name":"Department of Pharmacology and Toxicology College of Pharmacy, The University of Arizona Tucson Arizona"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Ryan D.","family":"Canatsey","sequence":"additional","affiliation":[{"name":"Southwest Environmental Health Sciences Center College of Pharmacy, The University of Arizona Tucson Arizona"},{"name":"Department of Pharmacology and Toxicology College of Pharmacy, The University of Arizona Tucson Arizona"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Terrence J.","family":"Monks","sequence":"additional","affiliation":[{"name":"Southwest Environmental Health Sciences Center College of Pharmacy, The University of Arizona Tucson Arizona"},{"name":"Department of Pharmacology and Toxicology College of Pharmacy, The University of Arizona Tucson Arizona"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Serrine S.","family":"Lau","sequence":"additional","affiliation":[{"name":"Southwest Environmental Health Sciences Center College of Pharmacy, The University of Arizona Tucson Arizona"},{"name":"Department of Pharmacology and Toxicology College of Pharmacy, The University of Arizona Tucson 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