Snyder L A, Saunders N J, Shafer W M
Department of Microbiology and Immunology, Emory University School of Medicine, Atlanta, Georgia 30322, USA.
J Bacteriol. 2001 Feb;183(4):1233-41. doi: 10.1128/JB.183.4.1233-1241.2001.
A cluster of 18 open reading frames (ORFs), 15 of which are homologous to genes involved in division and cell wall synthesis, has been identified in Neisseria gonorrhoeae and Neisseria meningitidis. The three additional ORFs, internal to the dcw cluster, are not homologous to dcw-related genes present in other bacterial species. Analysis of the N. meningitidis strain MC58 genome for foreign DNA suggests that these additional ORFs have not been acquired by recent horizontal exchange, indicating that they are a long-standing, integral part of the neisserial dcw gene cluster. Reverse transcription-PCR analysis of RNA extracted from N. gonorrhoeae strain FA19 confirmed that all three ORFs are transcribed in gonococci. One of these ORFs (dca, for division cluster competence associated), located between murE and murF, was studied in detail and found to be essential for competence in the gonococcal but not in the meningococcal strains tested. Computer analysis predicts that dca encodes an inner membrane protein similar to hypothetical proteins produced by other gram-negative bacteria. In some meningococcal strains dca is prematurely terminated following a homopolymeric tract of G's, the length of which differs between isolates of N. meningitidis, suggesting that dca is phase variable in this species. A deletion and insertional mutation was made in the dca gene of N. gonorrhoeae strain FA19 and N. meningitidis strain NMB. This mutation abrogated the ability of the gonococci to be transformed with chromosomal DNA. Thus, we conclude that the dca-encoded gene product is an essential competence factor for gonococci.
在淋病奈瑟菌和脑膜炎奈瑟菌中已鉴定出一组18个开放阅读框(ORF),其中15个与参与分裂和细胞壁合成的基因同源。dcw基因簇内部的另外三个ORF与其他细菌物种中存在的dcw相关基因不同源。对脑膜炎奈瑟菌菌株MC58基因组中的外源DNA分析表明,这些额外的ORF并非通过近期水平交换获得,这表明它们是奈瑟菌dcw基因簇长期以来不可或缺的一部分。对从淋病奈瑟菌菌株FA19中提取的RNA进行逆转录PCR分析证实,所有这三个ORF在淋球菌中均有转录。其中一个ORF(dca,与分裂簇能力相关)位于murE和murF之间,已进行了详细研究,发现它对所测试的淋球菌菌株的感受态至关重要,但对脑膜炎球菌菌株则不然。计算机分析预测,dca编码一种内膜蛋白,类似于其他革兰氏阴性细菌产生的假设蛋白。在一些脑膜炎球菌菌株中,dca在一段G的同聚物序列后过早终止,其长度在脑膜炎奈瑟菌分离株之间有所不同,这表明dca在该物种中是相位可变的。在淋病奈瑟菌菌株FA19和脑膜炎奈瑟菌菌株NMB的dca基因中进行了缺失和插入突变。这种突变消除了淋球菌用染色体DNA进行转化的能力。因此,我们得出结论,dca编码的基因产物是淋球菌必不可少的感受态因子。