Chen Y Y, Weaver C A, Mendelsohn D R, Burne R A
Center for Oral Biology, School of Medicine and Dentistry, University of Rochester, Rochester, New York 14642, USA.
J Bacteriol. 1998 Nov;180(21):5769-75. doi: 10.1128/JB.180.21.5769-5775.1998.
The Streptococcus salivarius 57.I ure cluster was organized as an operon, beginning with ureI, followed by ureABC (structural genes) and ureEFGD (accessory genes). Northern analyses revealed transcripts encompassing structural genes and transcripts containing the entire operon. A sigma70-like promoter could be mapped 5' to ureI (PureI) by primer extension analysis. The intensity of the signal increased when cells were grown at an acidic pH and was further enhanced by excess carbohydrate. To determine the function(s) of two inverted repeats located 5' to PureI, transcriptional fusions of the full-length promoter region (PureI), or a deletion derivative (PureIDelta100), and a promoterless chloramphenicol acetyltransferase (CAT) gene were constructed and integrated into the chromosome to generate strains PureICAT and PureIDelta100CAT, respectively. CAT specific activities of PureICAT were repressed at pH 7.0 and induced at pH 5.5 and by excess carbohydrate. In PureIDelta100CAT, CAT activity was 60-fold higher than in PureICAT at pH 7.0 and pH induction was nearly eliminated, indicating that expression was negatively regulated. Thus, it was concluded that PureI was the predominant, regulated promoter and that regulation was governed by a mechanism differing markedly from other known mechanisms for bacterial urease expression.
唾液链球菌57.I脲簇被组织成一个操纵子,从ureI开始,接着是ureABC(结构基因)和ureEFGD(辅助基因)。Northern分析揭示了包含结构基因的转录本以及包含整个操纵子的转录本。通过引物延伸分析可将一个类似sigma70的启动子定位到ureI的5'端(PureI)。当细胞在酸性pH下生长时,信号强度增加,并且过量碳水化合物会进一步增强该信号。为了确定位于PureI 5'端的两个反向重复序列的功能,构建了全长启动子区域(PureI)或缺失衍生物(PureIDelta100)与无启动子氯霉素乙酰转移酶(CAT)基因的转录融合体,并将其整合到染色体中,分别产生菌株PureICAT和PureIDelta100CAT。PureICAT的CAT比活性在pH 7.0时受到抑制,在pH 5.5时以及在过量碳水化合物存在下被诱导。在PureIDelta100CAT中,在pH 7.0时CAT活性比在PureICAT中高60倍,并且pH诱导几乎消除,表明表达受到负调控。因此,得出的结论是PureI是主要的、受调控的启动子,并且调控是由一种与细菌脲酶表达的其他已知机制明显不同的机制所控制。