Ubc9 fusion-directed SUMOylation (UFDS): a method to analyze function of protein SUMOylation. Export

@article{citeulike:1130546, abstract = {Although small ubiquitin-like modifier (SUMO) is conjugated to proteins involved in diverse cellular processes, the functional analysis of SUMOylated proteins is often hampered by low levels of specific SUMOylated proteins in the cell. Here we describe a SUMO-conjugating enzyme (Ubc9) fusion-directed SUMOylation (UFDS) system, which allows efficient and selective in vivo SUMOylation of proteins. Although SUMOylation of overexpressed p53 and STAT1 was difficult to detect in HEK293 cells, up to 40\% of p53 and STAT1 were conjugated with endogenous SUMO when fused to Ubc9. We verified the specificity of UFDS using SUMOylation-site mutants and showed that the method is not dependent on SUMO ligases. Using UFDS we demonstrated that SUMOylation of STAT1 inhibits its phosphorylation at Tyr701 and discovered p53 multi-SUMOylation in vivo. We propose that UFDS will be useful for the analysis of function of SUMOylation in protein interactions, subcellular localization as well as enzymatic activity.}, author = {Jakobs, Astrid and Koehnke, Jesko and Himstedt, Fabian and Funk, Martin and Korn, Bernhard and Gaestel, Matthias and Niedenthal, Rainer}, citeulike-article-id = {1130546}, citeulike-linkout-0 = {http://dx.doi.org/10.1038/nmeth1006}, citeulike-linkout-1 = {http://view.ncbi.nlm.nih.gov/pubmed/17277783}, citeulike-linkout-2 = {http://www.hubmed.org/display.cgi?uids=17277783}, day = {04}, doi = {10.1038/nmeth1006}, issn = {1548-7091}, journal = {Nature methods}, keywords = {methods, outside}, month = {March}, number = {3}, pages = {245--250}, posted-at = {2007-04-04 00:09:36}, priority = {3}, publisher = {Nature Publishing Group}, title = {Ubc9 fusion-directed SUMOylation (UFDS): a method to analyze function of protein SUMOylation.}, url = {http://dx.doi.org/10.1038/nmeth1006}, volume = {4}, year = {2007} }

TY - JOUR ID - citeulike:1130546 L3 - citeulike-article-id:1130546 N2 - Although small ubiquitin-like modifier (SUMO) is conjugated to proteins involved in diverse cellular processes, the functional analysis of SUMOylated proteins is often hampered by low levels of specific SUMOylated proteins in the cell. Here we describe a SUMO-conjugating enzyme (Ubc9) fusion-directed SUMOylation (UFDS) system, which allows efficient and selective in vivo SUMOylation of proteins. Although SUMOylation of overexpressed p53 and STAT1 was difficult to detect in HEK293 cells, up to 40% of p53 and STAT1 were conjugated with endogenous SUMO when fused to Ubc9. We verified the specificity of UFDS using SUMOylation-site mutants and showed that the method is not dependent on SUMO ligases. Using UFDS we demonstrated that SUMOylation of STAT1 inhibits its phosphorylation at Tyr701 and discovered p53 multi-SUMOylation in vivo. We propose that UFDS will be useful for the analysis of function of SUMOylation in protein interactions, subcellular localization as well as enzymatic activity. IS - 3 JF - Nature methods SN - 1548-7091 EP - 250 TI - Ubc9 fusion-directed SUMOylation (UFDS): a method to analyze function of protein SUMOylation. PB - Nature Publishing Group VL - 4 SP - 245 KW - methods KW - outside AU - Jakobs, Astrid AU - Koehnke, Jesko AU - Himstedt, Fabian AU - Funk, Martin AU - Korn, Bernhard AU - Gaestel, Matthias AU - Niedenthal, Rainer PY - 2007/03/04/ UR - http://dx.doi.org/10.1038/nmeth1006 ER -