title: Oncogenic KIT mutations in different exons lead to specific changes in melanocyte phospho-proteome.
abstract: Mutations in the proto-oncogene c-KIT (KIT) are found in several cancers, and the site of these mutations differs markedly between cancer types. We used site directed mutagenesis to induce KIT(559), KIT(642) and KIT(816) mutations in primary human melanocytes (PHM) and we investigated the impact of each mutation on KIT function. We studied canonical KIT-signaling pathways by immunoblotting, and we used stable isotope labeling by amino acids in cell culture (SILAC) and kinase prediction models to identify kinases differently activated in respective mutants. We validated our results with the analysis of phosphorylation levels of selected substrates for each kinase. We concluded that CK1 ε and δ are more active in cell clones harboring KIT(559) and KIT(642) mutations, whereas PAK4 is more active in clones with KIT(816) mutation. Our findings might help to develop further therapeutic options for tumors with specific KIT mutations in different domains.
  
  
  
  Different types of cancers harbor mutations in the oncogene KIT. The use of small molecules inhibitors directly targeting KIT had a limited success in the treatment of patients with KIT mutant cancers. Our study describes specific phospho-proteome changes due to different KIT mutations, and provides targets of further therapeutic options.
authors.affiliation: University of California San Francisco, Department of Dermatology, Mt. Zion Cancer Research Center, 2340 Sutter Street N461, 94115 San Francisco, USA; Department of Medical Sciences, Section of Dermatology, University of Turin, Italy. Electronic address: martina.sanlorenzo@hotmail.it.
authors.forename: M
authors.initials: M
authors.lastname: Sanlorenzo
authors.affiliation: University of California San Francisco, Department of Dermatology, Mt. Zion Cancer Research Center, 2340 Sutter Street N461, 94115 San Francisco, USA; The Rudolfstiftung Hospital, Academic Teaching Hospital, Medical University Vienna, Department of Dermatology, Juchgasse 25, 1030 Vienna, Austria; School of Medicine, Sigmund Freud University Vienna, Austria.
authors.forename: I
authors.initials: I
authors.lastname: Vujic
authors.affiliation: University of California San Francisco, Department of Dermatology, Mt. Zion Cancer Research Center, 2340 Sutter Street N461, 94115 San Francisco, USA; The Rudolfstiftung Hospital, Academic Teaching Hospital, Medical University Vienna, Department of Dermatology, Juchgasse 25, 1030 Vienna, Austria.
authors.forename: C
authors.initials: C
authors.lastname: Posch
authors.affiliation: University of California San Francisco, Department of Dermatology, Mt. Zion Cancer Research Center, 2340 Sutter Street N461, 94115 San Francisco, USA.
authors.forename: J
authors.initials: J
authors.lastname: Ma
authors.affiliation: University of California San Francisco, Department of Dermatology, Mt. Zion Cancer Research Center, 2340 Sutter Street N461, 94115 San Francisco, USA.
authors.forename: K
authors.initials: K
authors.lastname: Lin
authors.affiliation: University of California San Francisco, Department of Dermatology, Mt. Zion Cancer Research Center, 2340 Sutter Street N461, 94115 San Francisco, USA.
authors.forename: K
authors.initials: K
authors.lastname: Lai
authors.affiliation: University of California San Francisco, Department of Dermatology, Mt. Zion Cancer Research Center, 2340 Sutter Street N461, 94115 San Francisco, USA.
authors.forename: D
authors.initials: D
authors.lastname: Lee
authors.affiliation: University of California San Francisco, Department of Dermatology, Mt. Zion Cancer Research Center, 2340 Sutter Street N461, 94115 San Francisco, USA.
authors.forename: M
authors.initials: M
authors.lastname: Vujic
authors.affiliation: University of California San Francisco, School of Pharmacy, Pharmaceutical Chemistry, 600 16th Street, 94158 San Francisco, USA.
authors.forename: J A
authors.initials: JA
authors.lastname: Oses-Prieto
authors.affiliation: University of California San Francisco, School of Pharmacy, Pharmaceutical Chemistry, 600 16th Street, 94158 San Francisco, USA.
authors.forename: S
authors.initials: S
authors.lastname: Chand
authors.affiliation: Hospital Universitario 12 de Octubre, Madrid, Spain.
authors.forename: J L
authors.initials: JL
authors.lastname: Rodriguez-Peralto
authors.affiliation: University of California San Francisco, School of Pharmacy, Pharmaceutical Chemistry, 600 16th Street, 94158 San Francisco, USA.
authors.forename: A
authors.initials: A
authors.lastname: Burlingame
authors.affiliation: University of California San Francisco, Department of Dermatology, Mt. Zion Cancer Research Center, 2340 Sutter Street N461, 94115 San Francisco, USA.
authors.forename: S
authors.initials: S
authors.lastname: Ortiz-Urda
mesh_terms: D047388:Casein Kinases / Q000378:metabolism; D002478:Cells, Cultured; D005091:Exons; D006801:Humans; D008544:Melanocytes / Q000737:chemistry* / Q000378:metabolism; D058990:Molecular Targeted Therapy; D009154:Mutation; D009369:Neoplasms / Q000235:genetics; D010750:Phosphoproteins / Q000378:metabolism; D010766:Phosphorylation; D020543:Proteome / Q000378:metabolism*; D000090063:Proto-Oncogene Mas; D019009:Proto-Oncogene Proteins c-kit / Q000235:genetics*; D015398:Signal Transduction; D054462:p21-Activated Kinases / Q000378:metabolism
chemical_list: C000715470:MAS1 protein, human; D010750:Phosphoproteins; D020543:Proteome; D000090063:Proto-Oncogene Mas; C116367:PAK4 protein, human; D019009:Proto-Oncogene Proteins c-kit; D047388:Casein Kinases; D054462:p21-Activated Kinases
keywords: Kinases prediction; SILAC