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枯草芽孢杆菌的Dra-nupC-pdp操纵子:核苷酸序列、脱氧核苷诱导作用以及由deoR编码的DeoR阻遏蛋白的转录调控

Dra-nupC-pdp operon of Bacillus subtilis: nucleotide sequence, induction by deoxyribonucleosides, and transcriptional regulation by the deoR-encoded DeoR repressor protein.

作者信息

Saxild H H, Andersen L N, Hammer K

机构信息

Department of Microbiology, Technical University of Denmark, Lyngby, Denmark.

出版信息

J Bacteriol. 1996 Jan;178(2):424-34. doi: 10.1128/jb.178.2.424-434.1996.

Abstract

The genes encoding deoxyriboaldolase (dra), nucleoside uptake protein (nupC), and pyrimidine nucleoside sequences were determined. Sequence analysis showed that the genes were localized immediately downstream of the hut operon. Insertional gene disruption studies indicated that the three genes constitute an operon with the gene order dra-nupC-pdp. A promoter mapping immediately upstream of the dra gene was identified, and downstream of the pdp gene the nucleotide sequence indicated the existence of a factor-independent transcription terminator structure. In wild-type cells growing in succinate minimal medium, the pyrimidine nucleoside phosphorylase and deoxyriboaldolase levels were five- to eightfold higher in the presence of thymidine and fourfold higher in the presence of deoxyadenosine. By the use of lacZ fusions, the regulation was found to be at the level of transcription. The operon expression was subject to glucose repression. Upstream of the dra gene an open reading frame of 313 amino acids was identified. Inactivation of this gene led to an approximately 10-fold increase in the levels of deoxyriboaldolase and pyrimidine nucleoside phosphorylase, and no further induction was seen upon the addition of deoxyribonucleosides. The upstream gene most likely encodes the regulator for the dra-nupC-pdp operon and was designated deoR (stands for deoxyribonucleoside regulator).

摘要

确定了编码脱氧核糖醛缩酶(dra)、核苷摄取蛋白(nupC)和嘧啶核苷序列的基因。序列分析表明,这些基因位于hut操纵子的紧邻下游。插入基因破坏研究表明,这三个基因构成一个操纵子,基因顺序为dra - nupC - pdp。在dra基因紧邻上游确定了一个启动子图谱,在pdp基因下游,核苷酸序列表明存在一个不依赖因子的转录终止子结构。在琥珀酸基本培养基中生长的野生型细胞中,在胸苷存在下嘧啶核苷磷酸化酶和脱氧核糖醛缩酶水平高出五到八倍,在脱氧腺苷存在下高出四倍。通过使用lacZ融合,发现这种调节发生在转录水平。操纵子表达受到葡萄糖阻遏。在dra基因上游鉴定出一个由313个氨基酸组成的开放阅读框。该基因的失活导致脱氧核糖醛缩酶和嘧啶核苷磷酸化酶水平增加约10倍,添加脱氧核糖核苷后未观察到进一步的诱导。上游基因很可能编码dra - nupC - pdp操纵子的调节因子,被命名为deoR(代表脱氧核糖核苷调节因子)。

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