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原核生物金属硫蛋白基因座的分离及痕量金属离子对转录调控的分析。

Isolation of a prokaryotic metallothionein locus and analysis of transcriptional control by trace metal ions.

作者信息

Huckle J W, Morby A P, Turner J S, Robinson N J

机构信息

Department of Biological Sciences, University of Durham, UK.

出版信息

Mol Microbiol. 1993 Jan;7(2):177-87. doi: 10.1111/j.1365-2958.1993.tb01109.x.

DOI:10.1111/j.1365-2958.1993.tb01109.x
PMID:8446025
Abstract

In eukaryotes, metallothioneins (MTs) are involved in cellular responses to elevated concentrations of certain metal ions. We report the isolation and analysis of a prokaryotic MT locus from Synechococcus PCC 7942. The MT locus (smt) includes smtA, which encodes a class II MT, and a divergently transcribed gene, smtB. The sites of transcription initiation of both genes have been mapped and features within the smt operator-promoter region identified. Elevated concentrations of the ionic species of Cd, Co, Cr, Cu, Hg, Ni, Pb and Zn elicited an increase in the abundance of smtA transcripts. There was no detectable effect of elevated metal (Cd) on smtA transcript stability. Sequences upstream of smtA, fused to a promoterless lacZ gene, conferred metal-dependent beta-galactosidase activity in Synechococcus PCC 7942 (strain R2-PIM8). At maximum permissive concentrations, Zn was the most potent elicitor in vivo, followed by Cu and Cd with slight induction by Co and Ni. The deduced SmtB polypeptide has similarity to the ArsR and CadC proteins involved in resistance to arsenate/arsenite/antimonite and to Cd, contains a predicted helix-turn-helix DNA-binding motif and is shown to be a repressor of transcription from the smtA operator-promoter.

摘要

在真核生物中,金属硫蛋白(MTs)参与细胞对某些金属离子浓度升高的反应。我们报告了从聚球藻PCC 7942中分离和分析原核MT基因座的情况。MT基因座(smt)包括编码II类MT的smtA和一个反向转录的基因smtB。已确定了这两个基因的转录起始位点,并鉴定了smt操纵子 - 启动子区域内的特征。Cd、Co、Cr、Cu、Hg、Ni、Pb和Zn的离子种类浓度升高会导致smtA转录本丰度增加。金属(Cd)浓度升高对smtA转录本稳定性没有可检测到的影响。与无启动子的lacZ基因融合的smtA上游序列在聚球藻PCC 7942(菌株R2 - PIM8)中赋予了金属依赖性β - 半乳糖苷酶活性。在最大允许浓度下,Zn是体内最有效的诱导剂,其次是Cu和Cd,Co和Ni有轻微诱导作用。推导的SmtB多肽与参与抗砷酸盐/亚砷酸盐/锑酸盐和抗Cd的ArsR和CadC蛋白相似,含有预测的螺旋 - 转角 - 螺旋DNA结合基序,并被证明是smtA操纵子 - 启动子转录的阻遏物。

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