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rpbA控制费氏藻中组成型藻蓝蛋白基因集的转录。

rpbA controls transcription of the constitutive phycocyanin gene set in Fremyella diplosiphon.

作者信息

Kahn K, Schaefer M R

机构信息

Division of Molecular Biology and Biochemistry, School of Biological Sciences, University of Missouri-Kansas City, 64110, USA.

出版信息

J Bacteriol. 1997 Dec;179(24):7695-704. doi: 10.1128/jb.179.24.7695-7704.1997.

Abstract

Three gene sets encode alpha and beta subunits of the phycobiliprotein phycocyanin (PC) in the filamentous cyanobacterium Fremyella diplosiphon. The cpcB1A1 set (encodes PC1) is constitutively expressed, whereas the cpcB2A2 set (encodes PC2) is expressed only in red light and the cpcB3A3 set (encodes PC3) is expressed only during sulfur-limited growth. Primary pigment mutant strain FdBM1 is characterized by elevated levels of PC. DNA hybridization analysis showed that like many pigment mutants in our strain collection, strain FdBM1 harbors an extra genomic copy of endogenous transposon Tn5469. By direct cloning from FdBM1 genomic DNA, the extra copy of Tn5469 was localized to an open reading frame, which we have designated the rpbA gene. Complementation experiments correlated rpbA activity to the phenotype of strain FdBM1. The predicted RpbA protein contains two regions resembling the characterized helix-turn-helix motif which is involved in DNA recognition by many bacterial and phage transcription regulator proteins. RNA hybridization analysis showed that relative to the parental strain Fd33, the level of transcripts from cpcB1A1, but not cpcB2A2 or cpcB3A3, was significantly elevated in strain FdBM1. Introduction of the intact rpbA gene into strain FdBM1 restored the cpcB1A1 transcript level to that of strain Fd33. These results suggest that the rpbA gene product functions in controlling constitutive transcription from the cpcB1A1 gene set, possibly as a DNA-binding transcriptional repressor element.

摘要

在丝状蓝细菌双岐鱼腥藻(Fremyella diplosiphon)中,有三个基因集编码藻胆蛋白藻蓝蛋白(PC)的α和β亚基。cpcB1A1基因集(编码PC1)组成性表达,而cpcB2A2基因集(编码PC2)仅在红光下表达,cpcB3A3基因集(编码PC3)仅在硫限制生长期间表达。初级色素突变株FdBM1的特征是PC水平升高。DNA杂交分析表明,与我们菌株库中的许多色素突变体一样,菌株FdBM1含有内源性转座子Tn5469的一个额外基因组拷贝。通过从FdBM1基因组DNA直接克隆,Tn5469的额外拷贝被定位到一个开放阅读框,我们将其命名为rpbA基因。互补实验将rpbA活性与菌株FdBM1的表型相关联。预测的RpbA蛋白包含两个类似于特征性螺旋-转角-螺旋基序的区域,许多细菌和噬菌体转录调节蛋白通过该基序参与DNA识别。RNA杂交分析表明,相对于亲本菌株Fd33,cpcB1A1的转录本水平在菌株FdBM1中显著升高,而cpcB2A2或cpcB3A3的转录本水平没有升高。将完整的rpbA基因导入菌株FdBM1可使cpcB1A1转录本水平恢复到菌株Fd33的水平。这些结果表明,rpbA基因产物可能作为一种DNA结合转录抑制元件,在控制cpcB1A1基因集的组成性转录中发挥作用。

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