Yim H H, Brems R L, Villarejo M
Section of Microbiology, University of California, Davis 95616.
J Bacteriol. 1994 Jan;176(1):100-7. doi: 10.1128/jb.176.1.100-107.1994.
The osmY gene, which encodes a periplasmic protein with an apparent M(r) of 22,000, is induced by both osmotic and growth phase signals. We demonstrate here that osmY expression is regulated at the level of transcription and that transcription initiates 242 nucleotides upstream of the osmY open reading frame. Relative to the transcriptional start site, 5' deletions up to -36 did not inhibit osmY expression. 3' deletions that extended into the untranslated leader region affected the overall level of osmY::lacZ expression but did not affect inducibility. 5' and 3' deletions that extended past the transcriptional start region essentially abolished osmY expression, suggesting that there is a single promoter region. A putative promoter was identified, and its -10 region, TATATT, closely resembles the sigma 70 consensus -10 sequence, TATAAT. However, we show that osmY is not absolutely dependent on a functional sigma 70 for its expression. Since osmY expression does require rpoS (R. Hengge-Aronis, R. Lange, N. Henneberg, and D. Fischer, J. Bacteriol. 175:259-265, 1993), which encodes a stationary-phase sigma factor, sigma S (K. Tanaka, Y. Takayanagi, N. Fujita, A. Ishihama, and H. Takahashi, Proc. Natl. Acad. Sci. USA 90:3511-3515, 1993), E sigma S may be the form of RNA polymerase responsible for transcription of osmY.
osmY基因编码一种表观分子量为22000的周质蛋白,该基因受渗透压和生长阶段信号诱导。我们在此证明,osmY的表达在转录水平受到调控,转录起始于osmY开放阅读框上游242个核苷酸处。相对于转录起始位点,5'端缺失至-36并不抑制osmY的表达。延伸至非翻译前导区的3'端缺失影响osmY::lacZ的整体表达水平,但不影响诱导性。延伸超过转录起始区的5'端和3'端缺失基本消除了osmY的表达,表明存在一个单一的启动子区域。鉴定出一个推定的启动子,其-10区TATATT与σ70共有-10序列TATAAT非常相似。然而,我们表明osmY的表达并非绝对依赖于功能性的σ70。由于osmY的表达确实需要rpoS(R. Hengge-Aronis、R. Lange、N. Henneberg和D. Fischer,《细菌学杂志》175:259-265,1993年),rpoS编码一种稳定期σ因子σS(K. Tanaka、Y. Takayanagi、N. Fujita、A. Ishihama和H. Takahashi,《美国国家科学院院刊》90:3511-3515,1993年),EσS可能是负责osmY转录的RNA聚合酶形式。