Department of Obstetrics and Gynecology, University of Michigan Medical School, Ann Arbor, MI 48109-0617, United States.
Mol Cell Endocrinol. 2010 Nov 25;329(1-2):26-32. doi: 10.1016/j.mce.2010.05.014. Epub 2010 Jun 4.
We have identified a specific LHR mRNA binding protein that selectively binds to the polypyrimidine-rich bipartite sequence in the coding region of the LHR mRNA and accelerates its degradation. This process has been shown to be one of the mechanisms that is responsible for the loss of the steady-state levels of LHR mRNA following the preovulatory LH surge or the down regulation of the receptor in response to the administration of a pharmacological dose of LH or hCG. The trans factor, designated as the LHR mRNA binding protein (LRBP), was purified and its identity was established as being mevalonate kinase, an enzyme involved in cholesterol biosynthesis. When mevalonate kinase expression was abolished by treating cultured luteal cells with 25-hydroxycholesterol, the ability to undergo LH-induced down regulation of LHR mRNA was completely abrogated. Examination of the crystal structure of mevalonate kinase coupled with mutagenesis of the critical residues in the catalytic site revealed that the catalytic site is in close proximity to the LHR mRNA binding site. Further studies revealed that mevalonate kinase causes LHR mRNA degradation by acting as a translational suppressor by forming an untranslatable ribonucleoprotein (RNP) complex which is then targeted for degradation. These studies show that LHR expression in the ovary is regulated by a post-transcriptional mechanism mediated by mevalonate kinase thereby linking LHR expression with cholesterol metabolism.
我们已经鉴定出一种特定的 LHR mRNA 结合蛋白,它能特异性地与 LHR mRNA 编码区富含嘧啶的二聚体序列结合,并加速其降解。这一过程是导致 LH 峰排卵前或对药理学剂量 LH 或 hCG 给药导致受体下调时 LHR mRNA 稳态水平丧失的机制之一。这种转位因子被指定为 LHR mRNA 结合蛋白 (LRBP),已被纯化,并确定其身份为甲羟戊酸激酶,这是一种参与胆固醇生物合成的酶。当用 25-羟胆固醇处理培养的黄体细胞以消除甲羟戊酸激酶的表达时,LH 诱导的 LHR mRNA 下调能力完全被消除。对甲羟戊酸激酶的晶体结构的检查以及对催化位点关键残基的突变表明,催化位点与 LHR mRNA 结合位点非常接近。进一步的研究表明,甲羟戊酸激酶通过形成无翻译活性的核糖核蛋白(RNP)复合物来充当翻译抑制物,从而导致 LHR mRNA 降解,然后该复合物被靶向降解。这些研究表明,卵巢中 LHR 的表达受甲羟戊酸激酶介导的转录后机制调控,从而将 LHR 的表达与胆固醇代谢联系起来。