Regulation of urokinase receptor expression by phosphoglycerate kinase.
Regulation of urokinase receptor expression by phosphoglycerate kinase.
复制标题
磷酸甘油酸激酶对尿激酶受体表达的调节。
DOI：
10.1165/rcmb.2003-0104oc
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发表时间：
2004
影响因子：
6.4
通讯作者：
Idell,Steven
中科院分区：
文献类型：
--
作者：
Shetty,Sreerama;Muniyappa,Harish;Halady,PrathapKS;Idell,Steven
转录后调控是真核生物基因表达通过顺式相互作用调控的主要机制，顺式相互作用作为信使RNA （mRNA）稳定性快速改变的信号。尿激酶型纤溶酶原激活物受体（uPAR） mRNA的调控涉及uPAR mRNA编码区序列与50kd uPAR mRNA结合蛋白的相互作用。我们从人支气管上皮细胞（Beas2B）中纯化了该蛋白，并鉴定为磷酸甘油酸激酶（PGK）。我们通过聚合酶链反应克隆了PGK cDNA，并通过凝胶迁移和西北印迹技术表达了特异结合uPAR mRNA编码区的重组PGK蛋白。我们还通过免疫沉淀证实了PGK蛋白与uPAR mRNA的直接相互作用。在uPAR过量产生的H157肺癌细胞中，PGK过表达导致细胞质uPAR mRNA和细胞表面uPAR蛋白表达降低。uPAR mRNA表达减少导致uPAR mRNA稳定性降低。转染PGK cDNA的H157细胞中，3h -胸苷的掺入和迁移出现下降。这些结果表明，PGK在转录后水平调控uPAR的表达。
Post-transcriptional regulation represents a major mechanism by which eukaryotic gene expression is regulated throughcis-transinteractions that serve as signals for rapid alterations of messenger RNA (mRNA) stability. Regulation of urokinase-type plasminogen activator receptor (uPAR) mRNA involves the interaction of a uPAR mRNA coding region sequence with a 50 kD uPAR mRNA binding protein. We purified this protein from human bronchial epithelial (Beas2B) cells and identified it as phosphoglycerate kinase (PGK). We cloned PGK cDNA by polymerase chain reaction and expressed the recombinant PGK protein, which specifically bound the uPAR mRNA coding region by gel mobility shift and Northwestern blotting. We also confirmed a direct interaction of PGK protein with uPAR mRNA by immunoprecipitation. Overexpression of PGK in uPAR-overproducing H157 lung carcinoma cells resulted in decreased cytoplasmic uPAR mRNA and cell surface uPAR protein expression. Reduced uPAR mRNA expression involved decreased stability of the uPAR mRNA. Decline in3H-thymidine incorporation and migration occurred in H157 cells transfected with PGK cDNA. These results demonstrate that PGK regulates uPAR expression at the post-transcriptional level.