Cui Tracy X, Lin Grace, LaPensee Christopher R, Calinescu Anda-Alexandra, Rathore Maanjot, Streeter Cale, Piwien-Pilipuk Graciela, Lanning Nathan, Jin Hui, Carter-Su Christin, Qin Zhaohui S, Schwartz Jessica
Department of Molecular and Integrative Physiology, University of Michigan Medical School, Ann Arbor, Michigan 48109-5622, USA.
Mol Endocrinol. 2011 Apr;25(4):681-93. doi: 10.1210/me.2010-0232. Epub 2011 Feb 3.
Regulation of c-Fos transcription by GH is mediated by CCAAT/enhancer binding protein β (C/EBPβ). This study examines the role of C/EBPβ in mediating GH activation of other early response genes, including Cyr61, Btg2, Socs3, Zfp36, and Socs1. C/EBPβ depletion using short hairpin RNA impaired responsiveness of these genes to GH, as seen for c-Fos. Rescue with wild-type C/EBPβ led to GH-dependent recruitment of the coactivator p300 to the c-Fos promoter. In contrast, rescue with C/EBPβ mutated at the ERK phosphorylation site at T188 failed to induce GH-dependent recruitment of p300, indicating that ERK-mediated phosphorylation of C/EBPβ at T188 is required for GH-induced recruitment of p300 to c-Fos. GH also induced the occupancy of phosphorylated C/EBPβ and p300 on Cyr61, Btg2, and Socs3 at predicted C/EBP-cAMP response element-binding protein motifs in their promoters. Consistent with a role for ERKs in GH-induced expression of these genes, treatment with U0126 to block ERK phosphorylation inhibited their GH-induced expression. In contrast, GH-dependent expression of Zfp36 and Socs1 was not inhibited by U0126. Thus, induction of multiple early response genes by GH in 3T3-F442A cells is mediated by C/EBPβ. A subset of these genes is regulated similarly to c-Fos, through a mechanism involving GH-stimulated ERK 1/2 activation, phosphorylation of C/EBPβ, and recruitment of p300. Overall, these studies suggest that C/EBPβ, like the signal transducer and activator of transcription proteins, regulates multiple genes in response to GH.
生长激素(GH)对c-Fos转录的调控是由CCAAT/增强子结合蛋白β(C/EBPβ)介导的。本研究探讨了C/EBPβ在介导GH对其他早期反应基因(包括Cyr61、Btg2、Socs3、Zfp36和Socs1)激活中的作用。使用短发夹RNA使C/EBPβ缺失会损害这些基因对GH的反应性,就像c-Fos一样。用野生型C/EBPβ进行挽救可导致共激活因子p300依赖于GH募集到c-Fos启动子上。相反,用在T188处ERK磷酸化位点发生突变的C/EBPβ进行挽救未能诱导p300依赖于GH的募集,这表明T188处ERK介导的C/EBPβ磷酸化是GH诱导p300募集到c-Fos所必需的。GH还诱导了磷酸化的C/EBPβ和p300在Cyr61、Btg2和Socs3启动子中预测的C/EBP - 环磷酸腺苷反应元件结合蛋白基序上的占据。与ERK在GH诱导这些基因表达中的作用一致,用U0126处理以阻断ERK磷酸化会抑制它们的GH诱导表达。相反,Zfp36和Socs1的GH依赖性表达不受U0126抑制。因此,GH在3T3 - F442A细胞中对多个早期反应基因的诱导是由C/EBPβ介导的。这些基因中的一部分与c-Fos的调控方式相似,通过一种涉及GH刺激的ERK 1/2激活、C/EBPβ磷酸化和p300募集的机制。总体而言,这些研究表明C/EBPβ与信号转导和转录激活蛋白一样,可响应GH调节多个基因。