RRC ID 50025
著者 Komiya M, Ito A, Endo M, Hiruma D, Hattori M, Saitoh H, Yoshida M, Ozawa T.
タイトル A genetic screen to discover SUMOylated proteins in living mammalian cells.
ジャーナル Sci Rep
Abstract Post-translational modification by the Small Ubiquitin-related Modifier (SUMO) is indispensable for diverse biological mechanisms. Although various attempts have been made to discover novel SUMO substrate proteins to unveil the roles of SUMOylation, the reversibility of SUMOylation, and the differences in the SUMOylation level still makes it difficult to explore infrequently-SUMOylated proteins in mammalian cells. Here, we developed a method to screen for mammalian SUMOylated proteins using the reconstitution of split fluorescent protein fragments in living mammalian cells. Briefly, the cells harboring cDNAs of SUMOylated proteins were identified by the reconstituted fluorescence emission and separated by cell sorting. The method successfully identified 36 unreported SUMO2-substrate candidates with distinct intracellular localizations and functions. Of the candidates, we found Atac2, a histone acetyltransferase, was SUMOylated at a lysine 408, and further modified by multiple SUMOs without isoform specificity. Because the present method is applicable to other SUMO isoforms and mammalian cell-types, it could contribute to a deeper understanding of the role of SUMOylation in various biological contexts.
巻・号 7(1)
ページ 17443
公開日 2017-12-12
DOI 10.1038/s41598-017-17450-7
PII 10.1038/s41598-017-17450-7
PMID 29234079
PMC PMC5727073
MeSH 3T3 Cells Animals Blotting, Western Brain / metabolism Genetic Techniques* HEK293 Cells HeLa Cells Humans Immunoprecipitation Mice Microscopy, Confocal / methods Microscopy, Fluorescence / methods Proteins / chemistry Proteins / genetics Proteins / metabolism* Sumoylation* / physiology
IF 3.998
引用数 1
リソース情報
遺伝子材料 pAP3neo cDNA Library Mouse 10T-half (RDB02903)