Yonezawa Hideo, Kuramitsu Howard K, Nakayama Shu-ichi, Mitobe Jiro, Motegi Mizuho, Nakao Ryoma, Watanabe Haruo, Senpuku Hidenobu
Department of Bacteriology, National Institute of Infectious Diseases, 1-23-1 Toyama, Shinjuku-ku, Tokyo 162-8640, Japan.
Antimicrob Agents Chemother. 2008 Aug;52(8):2742-9. doi: 10.1128/AAC.00235-08. Epub 2008 May 19.
The two-component lantibiotic Smb is produced by Streptococcus mutans GS5. In the present study, we identified seven strains of S. mutans containing the smb gene cluster. These strains could be classified into high- and low-level Smb producers relative to the levels of Smb production by indicator strains in vitro. This classification was dependent upon the transcription levels of the structural smbA and smbB genes. Sequence analysis upstream of smbA in the high- and low-level Smb-producing strains revealed differences at nucleotide position -46 relative to the smbA start codon. Interestingly, the transcription start site was present upstream of the point mutation, indicating that both groups of strains have the same promoter constructs and that the differential expression of smbA and smbB mRNA occurred subsequent to transcription initiation. In addition, smbA::lacZ fusion expression was higher when it was regulated by the sequences of strains with high-level Smb activity than when it was regulated by the comparable region from strains with low-level Smb activity. Taken together, we conclude that high- or low-level Smb expression is dependent on the presence of a G or a T nucleotide at position -46 relative to the smbA translational start site in S. mutans Smb producers.
双组分羊毛硫抗生素Smb由变形链球菌GS5产生。在本研究中,我们鉴定出七株含有smb基因簇的变形链球菌菌株。相对于指示菌株在体外的Smb产生水平,这些菌株可分为高水平和低水平Smb产生菌。这种分类取决于结构基因smbA和smbB的转录水平。对高水平和低水平Smb产生菌株中smbA上游的序列分析显示,相对于smbA起始密码子,在核苷酸位置-46处存在差异。有趣的是,转录起始位点位于点突变的上游,这表明两组菌株具有相同的启动子结构,并且smbA和smbB mRNA的差异表达发生在转录起始之后。此外,当smbA::lacZ融合表达由具有高水平Smb活性的菌株序列调控时,其表达高于由具有低水平Smb活性的菌株的可比区域调控时的表达。综上所述,我们得出结论,高水平或低水平的Smb表达取决于变形链球菌Smb产生菌中相对于smbA翻译起始位点的-46位核苷酸是G还是T。