3' untranslated region-mediated regulation of luteinizing hormone/human chorionic gonadotropin receptor expression.
3' untranslated region-mediated regulation of luteinizing hormone/human chorionic gonadotropin receptor expression.
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3非翻译区介导的黄体生成激素/人绒毛膜促性腺激素受体表达的调节。
DOI：
10.1021/bi961019a
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发表时间：
1996
期刊：
影响因子：
--
通讯作者：
Menon,KM
中科院分区：
文献类型：
--
作者：
Lu,DL;Menon,KM
已在大鼠卵巢中鉴定出多种促黄体生成素/人绒毛膜促性腺激素（LH/hCG）受体mRNA（6.7、4.4、2.6和1.8 kb）。本实验室以前克隆了一个3.5kb的cDNA，它对应于大鼠卵巢中主要的LH/hCG受体mRNA的6.7kb转录本的3'非翻译区（3' UTR）。为了确定3' UTR对受体表达的影响，我们构建了对应于LH/hCG受体或荧光素酶的开放阅读框的cDNA，加上这些构建体，其中加入了与短链相关的3' UTR。（4.4 kb）及长（6.7kb）LH/hCG受体转录物，并测量用大T抗原（293 T）转化的293细胞中的受体或荧光素酶表达。125 I-hCG配体结合分析显示，3' UTR通过转录后事件抑制受体表达。首先，与单独的开放阅读框相比，3'UTR缩短了放线菌素D阻滞细胞中受体mRNA的半衰期。第二，LH/hCG受体mRNA的长3' UTR与显著较少的核糖体相关。还测定了LH/hCG受体3'UTR对荧光素酶表达的影响。短的3' UTR增加荧光素酶活性，而长的3' UTR降低荧光素酶表达。因此，短3' UTR对受体和荧光素酶表达发挥相反的作用。然而，长3' UTR中的序列足以抑制293 T细胞中的受体和荧光素酶表达。
Multiple luteinizing hormone/human chorionic gonadotropin (LH/hCG) receptor mRNAs (6.7, 4.4, 2.6, and 1.8 kb) have been identified in the rat ovary. Our laboratory has previously cloned a 3.5 kb cDNA that corresponds to the 3‘ untranslated region (3‘ UTR) of the 6.7 kb transcript, the major LH/hCG receptor mRNA in rat ovary. In order to determine the effects of the 3‘ UTR on receptor expression, we have constructed cDNAs corresponding to the open reading frame of LH/hCG receptor or luciferase, plus these constructs with the addition of the 3‘ UTRs associated with the short (4.4 kb) and long (6.7 kb) LH/hCG receptor transcripts, and measured receptor or luciferase expression in 293 cells transformed with large T antigen (293T). Ligand binding analysis with125I-hCG revealed that the 3‘ UTR inhibited receptor expression, which occurs through posttranscriptional events. First, the 3‘ UTRs reduced receptor mRNA half-life in actinomycin D-arrested cells, as compared to the open reading frame alone. Second, LH/hCG receptor mRNAs with the long 3‘ UTR associated with significantly fewer ribosomes. The effect of the LH/hCG receptor 3‘ UTRs on luciferase expression was also determined. The short 3‘ UTR increased luciferase activity, whereas the long 3‘ UTR decreased luciferase expression. Thus, the short 3‘ UTR exerts opposite effects on receptor and luciferase expression. However, sequences in the long 3‘ UTR are sufficient to inhibit both receptor and luciferase expression in 293T cells.