Interaction between 3' untranslated region of calcitonin receptor messenger ribonucleic acid (RNA) and adenylate/uridylate (AU)-rich element binding proteins (AU-rich RNA-binding factor 1 and Hu antigen R).
Interaction between 3' untranslated region of calcitonin receptor messenger ribonucleic acid (RNA) and adenylate/uridylate (AU)-rich element binding proteins (AU-rich RNA-binding factor 1 and Hu antigen R).
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降钙素受体信使核糖核酸 (RNA) 3 非翻译区与富含腺苷酸/尿苷酸 (AU) 的元件结合蛋白（富含 AU 的 RNA 结合因子 1 和 Hu 抗原 R）之间的相互作用。
DOI：
10.1210/en.2003-0862
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发表时间：
2004
期刊：
影响因子：
4.8
通讯作者：
S. Katayamá
中科院分区：
文献类型：
--
作者：
S. Yasuda;S. Wada;Y. Arao;M. Kogawa;F. Kayama;S. Katayamá
我们先前在小鼠破骨细胞中的结果表明，降钙素（CT）改变CT受体（CTR）mRNA的稳定性。CTR mRNA转录物在3'非翻译区（3' UTR）中含有几个富含腺苷酸/尿苷酸（Au）的不稳定元件。当小鼠CTR mRNA的3 'UTR被[α-（32）P]-尿苷5-三磷酸标记时，在转录物和几种细胞提取物之间观察到相互作用，包括来自破骨细胞祖单核细胞/巨噬细胞细胞系RAW 264.7的那些。相互作用蛋白的分子量范围为约35至50 kDa，类似于富含AU的RNA结合因子1（AUF 1）和Hu抗原R（HuR）。放射性标记的3 'UTR转录物与40-kDa蛋白结合，其可以从用AUF 1 p40转染的细胞中提取。为了确认结合特异性，将含有具有血凝素标签的AUF 1 p40的pSG 5载体构建体瞬时转染到NIH 3 T3细胞中。将提取物与添加了poly（A）的CTR 3 'UTR孵育。使用抗血凝素抗体对反应混合物进行免疫沉淀，提取沉淀的mRNA种类，并使用oligo-dT引物进行逆转录。发现对CTR mRNA序列的3 'UTR具有特异性的PCR引物产生PCR信号。当转染突变的AUF 1 p40时，未观察到信号。以类似于AUF 1结合的方式，HuR也被发现与3 'UTR结合。从小鼠破骨细胞提取的RNA也发现AUF 1 p40和HuR的特异性结合。CT处理破骨细胞对AUF 1的表达没有显著影响，但降低了HuR及其mRNA的水平。通过在四环素调节系统的控制下表达含有或不含有CTR 3 'UTR的荧光素酶报告基因构建体，进一步测试CTR 3' UTR在mRNA稳定性中的作用。结果表明，3 'UTR的加入显著缩短了荧光素酶报告基因的mRNA半衰期。这些结果表明，AUF 1 p40，HuR和CTR mRNA转录本的3 'UTR可能参与CTR mRNA表达的转录后调节。
Our previous results in mouse osteoclasts suggested that calcitonin (CT) alters CT receptor (CTR) mRNA stability. The CTR mRNA transcript contains several adenylate/uridylate (AU)-rich destabilizing elements in the 3' untranslated region (3'UTR). When the 3'UTR of mouse CTR mRNA was labeled by [alpha-(32)P]-uridine 5-triphosphate, interactions were observed between the transcript and several cell extracts, including those from the osteoclast progenitor monocyte/macrophage cell line, RAW 264.7. The molecular masses of the interacting proteins ranged from approximately 35 to 50 kDa, similar to AU-rich RNA-binding factor 1 (AUF1) and Hu antigen R (HuR). Radiolabeled 3'UTR transcripts bound with a 40-kDa protein, which could be extracted from cells transfected with AUF1 p40. To confirm the binding specificity, a pSG5 vector construct, containing the AUF1 p40 with an hemagglutinin tag, was transiently transfected into NIH3T3 cells. The extracts were incubated with poly(A)-added CTR3'UTR. The reaction mixture was immunoprecipitated using an antihemagglutinin antibody and precipitated mRNA species were extracted and reverse transcribed using oligo-dT primers. It was found that PCR primers specific for the 3'UTR of CTR mRNA sequence generated a PCR signal. No signal was observed when mutated AUF1 p40 was transfected. In a manner similar to the AUF1 binding, HuR was also found to bind to the 3'UTR. Specific binding of AUF1 p40 and HuR was also found with RNA extracted from mouse osteoclasts. Treatment of osteoclasts with CT did not significantly affect the expression of AUF1 but decreased the levels of HuR and its mRNA. The role of CTR3'UTR in mRNA stability was further tested by expressing luciferase reporter constructs that did, or did not, contain the CTR3'UTR, under the control of the tetracycline-regulatory system. The results showed that the addition of 3'UTR considerably shortened the mRNA half-life of the luciferase reporter gene. These results suggest that AUF1 p40, HuR, and the 3'UTR of the CTR mRNA transcript could be involved in posttranscriptional regulation of CTR mRNA expression.
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DOI：
10.1172/jci117634
发表时间：
1995
期刊：
The Journal of clinical investigation
影响因子：
--
作者：
S. Takahashi;S. Goldring;M. S. Katz;S. Hilsenbeck;R. Williams;G. Roodman
通讯作者：
S. Takahashi;S. Goldring;M. S. Katz;S. Hilsenbeck;R. Williams;G. Roodman
破骨细胞谱系细胞中小鼠 C1a 降钙素受体的降钙素依赖性下调涉及转录机制。
影响因子：
4.8
作者：
Inoue,D;Shih,C;Galson,DL;Goldring,SR;Horne,WC;Baron,R
通讯作者：
Baron,R
DOI：
--
发表时间：
1993-10
期刊：
The Journal of Cell Biology
影响因子：
--
作者：
J. Wilhelm;R. Vale
通讯作者：
J. Wilhelm;R. Vale
下调受体 mRNA 的 β-肾上腺素激动剂上调选择性结合 β-肾上腺素受体 mRNA 的 M(r) 35,000 蛋白。
DOI：
--
发表时间：
1992
期刊：
The Journal of biological chemistry
影响因子：
--
作者：
Port,JD;Huang,LY;Malbon,CC
通讯作者：
Malbon,CC