dc.contributor.author | Werner, Eric D. | |
dc.contributor.author | Lee, Jongsoon | |
dc.contributor.author | Hansen, Lone | |
dc.contributor.author | Yuan, Minsheng | |
dc.contributor.author | Shoelson, Steven E. | |
dc.date.accessioned | 2019-10-17T05:19:02Z | |
dc.date.issued | 2004 | |
dc.identifier.citation | Werner, Eric D., Jongsoon Lee, Lone Hansen, Minsheng Yuan, and Steven E. Shoelson. 2004. “Insulin Resistance Due to Phosphorylation of Insulin Receptor Substrate-1 at Serine 302.” Journal of Biological Chemistry 279 (34). American Society for Biochemistry & Molecular Biology (ASBMB): 35298–305. doi:10.1074/jbc.m405203200. | |
dc.identifier.issn | 0021-9258 | |
dc.identifier.issn | 1083-351X | |
dc.identifier.uri | http://nrs.harvard.edu/urn-3:HUL.InstRepos:41555765 | * |
dc.description.abstract | Inhibitory serine phosphorylation is a potential molecular mechanism for insulin resistance. We have developed a new variant of the yeast two-hybrid method, referred to as disruptive yeast tri-hybrid (Y3H), to identify inhibitory kinases and sites of phosphorylation in insulin receptors (IR) and IR substrates, IRS-1. Using IR and IRS-1 as bait and prey, respectively, and c-Jun NH2-terminal kinase (JNK1) as the disruptor, we now show that phosphorylation of IRS-1 Ser-307, a previously identified site, is necessary but not sufficient for JNK1-mediated disruption of IR/IRS-1 binding. We further identify a new phosphorylation site, Ser-302, and show that this too is necessary for JNK1-mediated disruption. Seven additional kinases potentially linked to insulin resistance similarly block IR/IRS-1 binding in the disruptive Y3H, but through distinct Ser-302- and Ser-307-independent mechanisms. Phosphospecific antibodies that recognize sequences surrounding Ser(P)-302 or Ser(P)-307 were used to determine whether the sites were phosphorylated under relevant conditions. Phosphorylation was promoted at both sites in Fao hepatoma cells by reagents known to promote Ser/Thr phosphorylation, including the phorbol ester phorbol 12-myristate 13-acetate, anisomycin, calyculin A, and insulin. The antibodies further showed that Ser(P)-302 and Ser(P)-307 are increased in animal models of obesity and insulin resistance, including genetically obese ob/ob mice, diet-induced obesity, and upon induction of hyperinsulinemia. These findings demonstrate that phosphorylation at both Ser-302 and Ser-307 is necessary for JNK1-mediated inhibition of the IR/IRS-1 interaction and that Ser-302 and Ser-307 are phosphorylated in parallel in cultured cells and in vivo under conditions that lead to insulin resistance. | |
dc.language.iso | en_US | |
dc.publisher | American Society for Biochemistry and Molecular Biology | |
dash.license | LAA | |
dc.title | Insulin Resistance Due to Phosphorylation of Insulin Receptor Substrate-1 at Serine 302 | |
dc.type | Journal Article | |
dc.description.version | Version of Record | |
dc.relation.journal | The Journal of Biological Chemistry | |
dash.depositing.author | Lee, Jongsoon::1c41246e8408e28799a79b02710fcde3::600 | |
dc.date.available | 2019-10-17T05:19:02Z | |
dash.workflow.comments | 1Science Serial ID 106225 | |
dc.identifier.doi | 10.1074/jbc.M405203200 | |
dash.source.volume | 279;34 | |
dash.source.page | 35298-35305 | |