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新月柄杆菌钩操纵子区域中发育调控启动子的鉴定、核苷酸序列及调控

Identification, nucleotide sequence, and control of developmentally regulated promoters in the hook operon region of Caulobacter crescentus.

作者信息

Chen L S, Mullin D, Newton A

出版信息

Proc Natl Acad Sci U S A. 1986 May;83(9):2860-4. doi: 10.1073/pnas.83.9.2860.

Abstract

The major flagellar proteins, including the flagellins and the hook protein, are synthesized periodically in the Caulobacter crescentus cell cycle at the time of flagellum assembly. Although fla genes are regulated at the transcriptional level [Ohta, N., Chen, L.-S., Swanson, E. & Newton, A. (1985) J. Mol. Biol. 186, 107-115], the 5' regulatory regions of C. crescentus genes have not been identified. We describe here the results of nuclease S1 protection assays that map the 5' ends of mRNAs synthesized in vivo from transcription units II (hook operon) and II.1 of the hook gene cluster and locate the corresponding promoter regions PII and PII.1. The two promoters are regulated with different periodicities in the cell cycle and have different genetic requirements for expression. The failure to detect transcripts from either PI or PII in Escherichia coli suggests that developmentally regulated promoters of C. crescentus have different recognition sequences from those of E. coli. There is little nucleotide sequence homology between PII and PII.1. There are, however, three regions of homology between PII and the nucleotide sequence 5' to the 29-kDa-flagellin-related gene, and two of these are in regions of dyad symmetry. We discuss the possibility that DNA-protein interactions at homologous nucleotide sequences like those identified in PII are part of a regulatory gene cascade that participates in timing fla gene expression in the C. crescentus cell cycle.

摘要

包括鞭毛蛋白和钩蛋白在内的主要鞭毛蛋白,在新月柄杆菌细胞周期中鞭毛组装时周期性合成。尽管鞭毛蛋白基因在转录水平受到调控[太田,N.,陈,L.-S.,斯旺森,E. & 牛顿,A.(1985年)《分子生物学杂志》186卷,107 - 115页],但新月柄杆菌基因的5'调控区尚未确定。我们在此描述核酸酶S1保护试验的结果,该试验绘制了体内从转录单元II(钩操纵子)和钩基因簇的II.1合成的mRNA的5'末端图谱,并定位了相应的启动子区域PII和PII.1。这两个启动子在细胞周期中以不同的周期受到调控,且表达所需的遗传条件不同。在大肠杆菌中未能检测到来自PI或PII的转录本,这表明新月柄杆菌发育调控的启动子与大肠杆菌的启动子具有不同的识别序列。PII和PII.1之间几乎没有核苷酸序列同源性。然而,PII与29 kDa鞭毛蛋白相关基因5'端的核苷酸序列之间存在三个同源区域,其中两个位于二元对称区域。我们讨论了在PII中鉴定出的同源核苷酸序列处的DNA - 蛋白质相互作用可能是参与新月柄杆菌细胞周期中鞭毛蛋白基因表达定时的调控基因级联反应的一部分的可能性。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/cffc/323406/0a1c69fb0f64/pnas00313-0091-a.jpg

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