Prolactin-induced Jak2 phosphorylation of RUSH: a key element in Jak/RUSH signaling.

Prolactin-induced Jak2 phosphorylation of RUSH: a key element in Jak/RUSH signaling.
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DOI:
10.1016/j.mce.2010.05.010
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发表时间:
2010-08-30
影响因子:
4.1
通讯作者:
Chilton, Beverly S.
Chilton, Beverly S.
中科院分区:
医学2区
文献类型:
--
作者:
Helmer, Rebecca A.;Panchoo, Marlyn;Dertien, Janet S.;Bhakta, Suhani M.;Hewetson, Aveline;Chilton, Beverly S.

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Jak2/Stat 介导的催乳素信号传导最终导致 Stat5a-DNA 结合。然而,并非所有 Jak2 依赖性基因都有 Stat5 位点。抑制剂的 Western 分析表明,Jak2 是催乳素诱导的 RUSH 磷酸化的近端中间体。 HRE-H9 细胞的转染测定显示 RUSH 结合位点介导催乳素增强 RUSH 基因的黄体酮依赖性转录的能力。 Jak2抑制剂或靶向RUSH位点突变阻断了催乳素作用。 RUSH 与核提取物中的磷酸化 Jak2 进行免疫共沉淀。 Jak2抑制剂消除了HRE-H9细胞中磷酸-RUSH的核库,而不是RUSH的核内容物。核仁附属伙伴,例如通过对与 RUSH/GST-RING 免疫共沉淀的核蛋白进行 μLC/MS/MS 分析来鉴定核仁素。 RUSH 并不完全与纤维蛋白共定位于核仁。 MG-132(蛋白酶体抑制剂)无法阻断 Tyrene CR4 介导的磷酸-RUSH 减少,并且不会促进核仁中 RUSH 的积累。这些研究证实了 RUSH 的催乳素依赖性 Jak2 磷酸化,并提供了对核相互作用的 RUSH 网络的功能影响。
Jak2/Stat-mediated prolactin signaling culminates in Stat5a-DNA-binding. However, not all Jak2-dependent genes have Stat5 sites. Western analysis with inhibitors showed Jak2 is a proximal intermediate in prolactin-induced RUSH phosphorylation. Transfection assays with HRE-H9 cells showed the RUSH-binding site mediated the ability of prolactin to augment progesterone-dependent transcription of the RUSH gene. Jak2 inhibitors or targeted RUSH-site mutation blocked the prolactin effect. RUSH co-immunoprecipitated with phospho-Jak2 from nuclear extracts. Jak2 inhibitors abolished the nuclear pool of phospho-RUSH not the nuclear content of RUSH in HRE-H9 cells. Nucleolar-affiliated partners, e.g. nucleolin, were identified by μLC/MS/MS analysis of nuclear proteins that co-immunoprecipitated with RUSH/GST-RING. RUSH did not exclusively co-localize with fibrillarin to the nucleolus. MG-132 (proteasomal inhibitor) failed to block Tyrene CR4-mediated decrease in phospho-RUSH, and did not promote RUSH accumulation in the nucleolus. These studies authenticate prolactin-dependent Jak2 phosphorylation of RUSH, and provide functional implications on the RUSH network of nuclear interactions.
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