Perego M, Spiegelman G B, Hoch J A
Division of Cellular Biology, Research Institute of Scripps Clinic, La Jolla, California 92037.
Mol Microbiol. 1988 Nov;2(6):689-99. doi: 10.1111/j.1365-2958.1988.tb00079.x.
Sporulation begins coincidentally with the expression of several stationary-phase-associated gene products during the transition state of a culture from exponential to stationary phase. Mutations in the stage 0 sporulation genes prevent the expression of these gene products in addition to blocking sporulation. Suppressor mutations in the abrB gene, in a spo0 background, restore stationary-phase-associated gene expression but not sporulation. The nature of the abrB gene product was investigated by isolating and sequencing the abrB gene. The abrB gene coded for a 96-amino-acid protein (molecular weight 10773) and contained a helix-turn-helix structure common to DNA binding proteins. Analysis of expression of the abrB gene using lacZ transcription fusions and direct measurement of mRNA content by hybridization showed that the spo0A gene repressed transcription of the abrB gene. Primer extension analysis of abrB gene mRNA revealed two initiation sites. The downstream site was dramatically repressed in spo0A+ strains, while the upstream site appeared not to be regulated by spo0A. Five abrB mutant alleles were cloned and sequenced. One mutation, abrB4, resided within the structural gene and continued to overexpress abrB messenger RNA from both promoters. A promoter mutation, abrB15, reduced transcription from the downstream promoter but not the upstream promoter. Thus, the phenotype of abrB mutations results from inactivation of the abrB gene product or by prevention of its overexpression. The results suggest that the abrB gene codes for a regulator which controls several genes whose products are normally produced during the transition phase between active growth and sporulation. The level of this regulator is, in turn, controlled by the spo0A gene. The pleiotropic phenotypes of spo0A mutants result from uncontrolled overexpression of the abrB regulator.
在培养物从指数生长期向稳定期转变的过渡阶段,芽孢形成与几种稳定期相关基因产物的表达同时开始。0期芽孢形成基因的突变除了阻断芽孢形成外,还会阻止这些基因产物的表达。在spo0背景下,abrB基因中的抑制突变可恢复稳定期相关基因的表达,但不能恢复芽孢形成。通过分离和测序abrB基因,研究了abrB基因产物的性质。abrB基因编码一种96个氨基酸的蛋白质(分子量10773),并含有DNA结合蛋白常见的螺旋-转角-螺旋结构。使用lacZ转录融合分析abrB基因的表达,并通过杂交直接测量mRNA含量,结果表明spo0A基因抑制abrB基因的转录。对abrB基因mRNA进行引物延伸分析,发现了两个起始位点。在spo0A+菌株中,下游位点受到显著抑制,而上游位点似乎不受spo0A调控。克隆并测序了五个abrB突变等位基因。一个突变体abrB4位于结构基因内,两个启动子均持续过度表达abrB信使RNA。一个启动子突变体abrB15减少了下游启动子的转录,但不影响上游启动子。因此,abrB突变的表型是由于abrB基因产物失活或其过度表达受阻所致。结果表明,abrB基因编码一种调节因子,该调节因子控制几个基因,这些基因的产物通常在活跃生长和芽孢形成之间的过渡阶段产生。而这种调节因子的水平又受spo0A基因的控制。spo0A突变体的多效表型是由于abrB调节因子的不受控制的过度表达所致。