A genetic screen to discover SUMOylated proteins in living mammalian cells

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...

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Autores principales:	Maki Komiya, Akihiro Ito, Mizuki Endo, Daisuke Hiruma, Mitsuru Hattori, Hisato Saitoh, Minoru Yoshida, Takeaki Ozawa
Formato:	article
Lenguaje:	EN
Publicado:	Nature Portfolio 2017
Materias:	Medicine R Science Q
Acceso en línea:	https://doaj.org/article/58242896948f4ac4bcd84defc6243b9c
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spelling	oai:doaj.org-article:58242896948f4ac4bcd84defc6243b9c2021-12-02T15:05:48ZA genetic screen to discover SUMOylated proteins in living mammalian cells10.1038/s41598-017-17450-72045-2322https://doaj.org/article/58242896948f4ac4bcd84defc6243b9c2017-12-01T00:00:00Zhttps://doi.org/10.1038/s41598-017-17450-7https://doaj.org/toc/2045-2322Abstract 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.Maki KomiyaAkihiro ItoMizuki EndoDaisuke HirumaMitsuru HattoriHisato SaitohMinoru YoshidaTakeaki OzawaNature PortfolioarticleMedicineRScienceQENScientific Reports, Vol 7, Iss 1, Pp 1-13 (2017)
institution	DOAJ
collection	DOAJ
language	EN
topic	Medicine R Science Q
spellingShingle	Medicine R Science Q Maki Komiya Akihiro Ito Mizuki Endo Daisuke Hiruma Mitsuru Hattori Hisato Saitoh Minoru Yoshida Takeaki Ozawa A genetic screen to discover SUMOylated proteins in living mammalian cells
description	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.
format	article
author	Maki Komiya Akihiro Ito Mizuki Endo Daisuke Hiruma Mitsuru Hattori Hisato Saitoh Minoru Yoshida Takeaki Ozawa
author_facet	Maki Komiya Akihiro Ito Mizuki Endo Daisuke Hiruma Mitsuru Hattori Hisato Saitoh Minoru Yoshida Takeaki Ozawa
author_sort	Maki Komiya
title	A genetic screen to discover SUMOylated proteins in living mammalian cells
title_short	A genetic screen to discover SUMOylated proteins in living mammalian cells
title_full	A genetic screen to discover SUMOylated proteins in living mammalian cells
title_fullStr	A genetic screen to discover SUMOylated proteins in living mammalian cells
title_full_unstemmed	A genetic screen to discover SUMOylated proteins in living mammalian cells
title_sort	genetic screen to discover sumoylated proteins in living mammalian cells
publisher	Nature Portfolio
publishDate	2017
url	https://doaj.org/article/58242896948f4ac4bcd84defc6243b9c
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A genetic screen to discover SUMOylated proteins in living mammalian cells

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