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Different interactomes for p70‐S6K1 and p54‐S6K2 revealed by proteomic analysis
Author(s) -
Pavan Isadora C. B.,
Yokoo Sami,
Granato Daniela C.,
Meneguello Letícia,
Carnielli Carolina M.,
Tavares Mariana R.,
Amaral Camila L.,
Freitas Lidia B.,
Paes Leme Adriana F.,
Luchessi Augusto D.,
Simabuco Fernando M.
Publication year - 2016
Publication title -
proteomics
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.26
H-Index - 167
eISSN - 1615-9861
pISSN - 1615-9853
DOI - 10.1002/pmic.201500249
Subject(s) - interactome , p70 s6 kinase 1 , biology , stable isotope labeling by amino acids in cell culture , alternative splicing , computational biology , pi3k/akt/mtor pathway , microbiology and biotechnology , proteomics , immunoprecipitation , rna splicing , ribosome biogenesis , protein–protein interaction , exon , signal transduction , ribosome , genetics , gene , rna
S6Ks are major effectors of the mTOR (mammalian target of rapamycin) pathway, signaling for increased protein synthesis and cell growth in response to insulin, AMP/ATP levels, and amino acids. Deregulation of this pathway has been related to disorders and diseases associated with metabolism, such as obesity, diabetes, and cancer. S6K family is composed of two main members, S6K1 and S6K2, which comprise different isoforms resulted from alternative splicing or alternative start codon use. Although important molecular functions have been associated with p70‐S6K1, the most extensively studied isoform, the S6K2 counterpart lacks information. In the present study, we performed immunoprecipitation assays followed by mass spectrometry (MS) analysis of FLAG‐tagged p70‐S6K1 and p54‐S6K2 interactomes, after expression in HEK293 cells. Protein lists were submitted to CRAPome (Contaminant Repository for Affinity Purification) and SAINT (Significance Analysis of INTeractome) analysis, which allowed the identification of high‐scoring interactions. By a comparative approach, p70‐S6K1 interacting proteins were predominantly related to “cytoskeleton” and “stress response,” whereas p54‐S6K2 interactome was more associated to “transcription,” “splicing,” and “ribosome biogenesis.” Moreover, we have found evidences for new targets or regulators of the S6K protein family, such as proteins NCL, NPM1, eIF2α, XRCC6, PARP1, and ILF2/ILF3 complex. This study provides new information about the interacting networks of S6Ks, which may contribute for future approaches to a better understanding of the mTOR/S6K pathway.