Rat liver glutathione S-transferases. DNA sequence analysis of a Yb2 cDNA clone and regulation of the Yb1 and Yb2 mRNAs by phenobarbital.
Rat liver glutathione S-transferases. DNA sequence analysis of a Yb2 cDNA clone and regulation of the Yb1 and Yb2 mRNAs by phenobarbital.
复制标题
大鼠肝脏谷胱甘肽 S-转移酶。
DOI：
10.1016/s0021-9258(19)57495-6
复制
发表时间：
1986
期刊：
影响因子：
--
通讯作者：
C. B. Pickett
中科院分区：
文献类型：
--
作者：
G. Ding;V. D. Ding;J. Rodkey;C. Bennett;A. Y. Lu;C. B. Pickett
我们已经构建了一个cDNA克隆，pGTA/C48，这是互补的大鼠肝谷胱甘肽S-转移酶Yb 2 mRNA。重组克隆pGTA/C48含有845个碱基对的cDNA插入片段，它与我们实验室先前描述的Yb 1 cDNA克隆pGTA/C44的核苷酸108-952重叠（Ding，G. J. - F.、Lu，中国茶条A. Y. H、Pickett，C. B。（1985）J.Biol.Chem.260，13268-13271）。在Yb 1和Yb 2 cDNA克隆的蛋白质编码区上有84%的核苷酸序列同源性，而3'非翻译区只有32%的同源性。Yb 2亚基的完整氨基酸序列已确定从pGTA/C48的DNA序列分析和谷胱甘肽S-转移酶Yb 1 Yb 2异二聚体的常规蛋白质序列分析的组合。Yb 2亚基由218个氨基酸组成，分子量为25，705，其氨基酸序列与Yb 1亚基的序列具有79%的同源性。我们利用3个大鼠肝谷胱甘肽S-转移酶cDNA克隆的不同3'非翻译区作为特异性探针，以确定苯巴比妥对Yb 1，Yb 2和Yc mRNA水平的影响。我们的研究结果清楚地表明，Yb 1和Yb 2的mRNA升高约5-6倍苯巴比妥管理，而Yc的mRNA仅适度升高，这种异生物质。最后，我们的数据表明，Yb 2亚基是由一个基因（S），这是从Yb 1基因（S）不同的编码，并提供了直接的证据，在大鼠中存在多个谷胱甘肽S-转移酶Yb基因。
We have constructed a cDNA clone, pGTA/C48, which is complementary to the rat liver glutathione S-transferase Yb2 mRNA. Recombinant clone pGTA/C48 contains a cDNA insert of 845 base pairs which overlaps nucleotides 108-952 of the Yb1 cDNA clone, pGTA/C44, described previously by our laboratory (Ding, G. J.-F., Lu, A. Y. H., and Pickett, C. B. (1985) J. Biol. Chem. 260, 13268-13271). Over the protein coding region of the Yb1 and Yb2 cDNA clones there is an 84% nucleotide sequence homology, whereas the 3‘ untranslated regions are only 32% homologous. The complete amino acid sequence of the Yb2 subunit has been determined from a combination of DNA sequence analysis of pGTA/C48 and conventional protein sequence analysis of the glutathione S-transferase Yb1 Yb2 heterodimer. The Yb2 subunit is comprised of 218 amino acids with a molecular weight of 25,705 and has an amino acid sequence which is 79% homologous to the sequence of the Yb1 subunit. We have utilized the divergent 3‘ untranslated regions of three rat liver glutathione S-transferase cDNA clones as specific probes to determine the effect of phenobarbital on the level of Yb1, Yb2, and Yc mRNAs. Our results clearly show that the Yb1 and Yb2 mRNAs are elevated approximately 5-6-fold by phenobarbital administration; whereas the Yc mRNA is only modestly elevated by this xenobiotic. Finally, our data suggest that the Yb2 subunit is encoded by a gene(s) which is distinct from the Yb1 gene(s) and provides direct evidence for the existence of multiple glutathione S-transferase Yb genes in the rat.