In vivo localization and identification of SUMOylated proteins in the brain of His6-HA-SUMO1 knock-in mice

SUMOylation, an essential posttranslational protein modification, is involved in many eukaryotic cellular signaling pathways. The identification of SUMOylated proteins is difficult, because SUMOylation sites in proteins are hard to predict, SUMOylated protein states are transient in vivo and labile...

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Main Authors: Tirard, Marilyn (Author) , Hsiao, He-Hsuan (Author) , Nikolov, Miroslav (Author) , Urlaub, Henning (Author) , Melchior, Frauke (Author) , Brose, Nils (Author)
Format: Article (Journal)
Language:English
Published: 2012
In: Proceedings of the National Academy of Sciences of the United States of America
Year: 2012, Volume: 109, Issue: 51, Pages: 21122-21127
ISSN:1091-6490
DOI:10.1073/pnas.1215366110
Online Access:Verlag, kostenfrei, Volltext: http://dx.doi.org/10.1073/pnas.1215366110
Verlag, kostenfrei, Volltext: http://www.pnas.org/content/109/51/21122
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Author Notes:Marilyn Tirard, He-Hsuan Hsiao, Miroslav Nikolov, Henning Urlaub, Frauke Melchior, and Nils Brose
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Summary:SUMOylation, an essential posttranslational protein modification, is involved in many eukaryotic cellular signaling pathways. The identification of SUMOylated proteins is difficult, because SUMOylation sites in proteins are hard to predict, SUMOylated protein states are transient in vivo and labile in vitro, only a small substrate fraction is SUMOylated in vivo, and identification tools for natively SUMOylated proteins are rare. To solve these problems, we generated knock-in mice expressing His6-HA-SUMO1. By anti-HA immunostaining, we show that SUMO1 conjugates in neurons are only detectable in nuclei and annulate lamellae. By anti-HA affinity purification, we identified several hundred candidate SUMO1 substrates, of which we validated Smchd1, Ctip2, TIF1γ, and Zbtb20 as novel substrates. The knock-in mouse represents an excellent mammalian model for studies on SUMO1 localization and screens for SUMO1 conjugates in vivo.
Item Description:Published first December 3, 2012
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Im Titel ist bei "His6-HA-SUMO1" die 6 tiefgestellt
Physical Description:Online Resource
ISSN:1091-6490
DOI:10.1073/pnas.1215366110