Margolis P S, Driks A, Losick R
Department of Cellular and Developmental Biology, Harvard University, Cambridge, Massachusetts 02138.
J Bacteriol. 1993 Jan;175(2):528-40. doi: 10.1128/jb.175.2.528-540.1993.
We have cloned and characterized the sporulation gene spoIIB from Bacillus subtilis. In extension of previous nucleotide sequence analysis, our results show that the order of genes in the vicinity of spoIIB is valS folC comC spoIIB orfA orfB mreB mreC mreD minC minD spoIVFA spoIVFB L20 orfX L24 spoOB obg pheB pheA. All 20 genes have the same orientation; the direction of transcription is from valS to pheA. We show that spoIIB is a 332-codon-long open reading frame whose transcription is under sporulation control. The deduced amino acid sequence of the spoIIB gene product, a 36-kDa polypeptide, is highly charged and contains a stretch of uncharged amino acids that could correspond to a transmembrane segment. Surprisingly, mutations in spoIIB, including an in vitro-constructed null mutation, cause only a mild impairment of spore formation in certain otherwise wild-type bacteria. However, when combined with mutations in another sporulation gene, spoVG, mutations in spoIIB cause a severe block in spore formation at the stage (stage II) of septum formation. (As with spoIIB mutations, mutations in spoVG cause little impairment in sporulation on their own.) The nature of the spoIIB spoVG mutant phenotype is discussed in terms of the events involved in the maturation of the sporulation septum and in the activation of sporulation transcription factors sigma F and sigma E.
我们已经克隆并鉴定了来自枯草芽孢杆菌的芽孢形成基因spoIIB。在先前核苷酸序列分析的基础上,我们的结果表明,spoIIB附近基因的顺序是valS folC comC spoIIB orfA orfB mreB mreC mreD minC minD spoIVFA spoIVFB L20 orfX L24 spoOB obg pheB pheA。所有20个基因具有相同的方向;转录方向是从valS到pheA。我们表明,spoIIB是一个332个密码子长的开放阅读框,其转录受芽孢形成控制。spoIIB基因产物是一种36 kDa的多肽,推导的氨基酸序列带电荷很高,并且包含一段可能对应于跨膜区段的不带电荷的氨基酸。令人惊讶的是,spoIIB中的突变,包括体外构建的无效突变,在某些其他方面为野生型的细菌中仅导致芽孢形成的轻微受损。然而,当与另一个芽孢形成基因spoVG中的突变结合时,spoIIB中的突变在隔膜形成阶段(阶段II)导致芽孢形成严重受阻。(与spoIIB突变一样,spoVG中的突变本身对芽孢形成几乎没有损害。)根据参与芽孢形成隔膜成熟以及芽孢形成转录因子sigma F和sigma E激活的事件,讨论了spoIIB spoVG突变体表型的性质。