Sabelnikov A G, Greenberg B, Lacks S A
Department of Biology, Brookhaven National Laboratory, Upton, NY 11973, USA.
J Mol Biol. 1995 Jul 7;250(2):144-55. doi: 10.1006/jmbi.1995.0366.
The genetic cassette encoding the DpnII restriction-modification system of Streptococcus pneumoniae gave transcription products of approximately 2.7 and 1.8 kilobases. The larger, mRNA1, covered both of the methylase genes, dpnM and dpnA, and the endonuclease gene dpnB; the smaller, mRNA2, covered only the dpnA and dpnB genes. Transcription of mRNA1 was shown to begin at the translation start site for dpnM, thereby producing an mRNA without any apparent ribosome-binding site for translation of the DpnM methylase. The promoter for mRNA1 was shown by base substitution and deletion analysis to consist of an extended -10 site, TaTGgTATAAT, with no required -35 site. A possible promoter further upstream with close matches to a -35 site and a nonextended -10 site was not used. A survey of 36 proven and putative promoters used by S. pneumoniae revealed that 61% of them contained the full -10 extension, although, other than the dpnM promoter, they matched at a -35 site, as well. It appears that, unlike those found in Escherichia coli, S. pneumoniae promoters frequently require an extended -10 site, and such a site can function naturally without a -35 site.
编码肺炎链球菌DpnII限制修饰系统的遗传盒产生了约2.7和1.8千碱基的转录产物。较大的mRNA1覆盖了甲基化酶基因dpnM和dpnA以及核酸内切酶基因dpnB;较小的mRNA2仅覆盖dpnA和dpnB基因。已证明mRNA1的转录起始于dpnM的翻译起始位点,从而产生一种没有任何明显核糖体结合位点用于DpnM甲基化酶翻译的mRNA。通过碱基置换和缺失分析表明,mRNA1的启动子由一个延伸的-10位点TaTGgTATAAT组成,不需要-35位点。上游一个与-35位点和非延伸-10位点紧密匹配的可能启动子未被使用。对肺炎链球菌使用的36个已证实和推测的启动子进行的调查显示,其中61%包含完整的-10延伸,尽管除了dpnM启动子外,它们在-35位点也匹配。看来,与在大肠杆菌中发现的启动子不同,肺炎链球菌的启动子经常需要一个延伸的-10位点,并且这样一个位点可以在没有-35位点的情况下自然发挥作用。