Lai H C, Li N, Weiss M J, Reddy C C, Tu C P
J Biol Chem. 1984 May 10;259(9):5536-42.
We have determined the nucleotide sequence of a cloned cDNA derived from liver poly(A) RNA of pentobarbital-treated rats encoding a glutathione S-transferase subunit. This cDNA clone pGTR261 contains one open reading frame of 222 amino acids, a complete 3' noncoding region, and 63 nucleotides in the 5' noncoding region. The cloned DNA hybridizes to rat poly(A) RNA in a tissue-specific fashion, with strong signals to liver and kidney poly(A) RNA(s) of approximately 1100 and approximately 1400 nucleotides in size but little or no hybridization to poly(A) RNAs from heart, lung, seminal vesicles, spleen, or testis under stringent conditions. Our sequence covers the cDNA sequence of pGST94 which contains a partial coding sequence for a liver glutathione S-transferase subunit of Ya size. Comparison of sequences with our earlier clone pGTR112 suggests that there are at least two mRNA species coding for two different subunits of the Ya (Mr = 25,600) subunit family with very limited amino acid substitutions mainly of conserved polarity. The divergent 3' noncoding sequences should be useful molecular probes in differentiating these two different but otherwise very similar subunits in induction and genomic structure analyses. Our results suggest that tissue-specific expression of the glutathione S-transferase subunits represented by the sequences of pGTR261 and pGTR112 may occur at or prior to the level of RNA processing.
我们已经确定了一个克隆cDNA的核苷酸序列,该cDNA来源于经戊巴比妥处理的大鼠肝脏多聚腺苷酸(poly(A))RNA,编码一种谷胱甘肽S-转移酶亚基。这个cDNA克隆pGTR261包含一个222个氨基酸的开放阅读框、一个完整的3'非编码区以及5'非编码区的63个核苷酸。克隆的DNA以组织特异性方式与大鼠多聚腺苷酸RNA杂交,在严格条件下,与大小约为1100和1400个核苷酸的肝脏和肾脏多聚腺苷酸RNA有强烈信号,但与来自心脏、肺、精囊、脾脏或睾丸的多聚腺苷酸RNA几乎没有杂交。我们的序列涵盖了pGST94的cDNA序列,pGST94包含一个Ya大小的肝脏谷胱甘肽S-转移酶亚基的部分编码序列。将该序列与我们早期的克隆pGTR112进行比较表明,至少有两种mRNA编码Ya(Mr = 25,600)亚基家族的两个不同亚基,氨基酸取代非常有限,主要是保守极性的取代。不同的3'非编码序列在诱导和基因组结构分析中区分这两个不同但其他方面非常相似的亚基时应是有用的分子探针。我们的结果表明,由pGTR261和pGTR112序列代表的谷胱甘肽S-转移酶亚基的组织特异性表达可能发生在RNA加工水平或之前。