Taha M K, So M, Seifert H S, Billyard E, Marchal C
Unité des Antigènes Bactériens, Institut Pasteur, Paris, France.
EMBO J. 1988 Dec 20;7(13):4367-78. doi: 10.1002/j.1460-2075.1988.tb03335.x.
We have identified two closely linked genes, pilA and pilB, which act in trans on the pilin promoter. pilA-pilB map downstream of expression loci pilE1 and opaE1 in the gonococcal chromosome. Subcloning data indicate that pilB acts negatively on the pilin promoter, and insertional inactivation of pilB results in hyperpiliated gonococci. A pilA clone activates the pilin promoter in Escherichia coli, and a pilA-/pilA+ heterodiploid gonococcus exhibits a P- phenotype. Our inability to obtain simple pilA- mutants strongly suggests that pilA is an essential gene in the gonococcus. In an in vitro coupled transcription/translation system, inserts spanning the pilA and pilB region direct the synthesis of two proteins of 40 and 58 kd. DNA sequence analysis shows that the pilA and pilB loci encode proteins of 38.6 kd (with a putative DNA binding domain) and 57.9 kd respectively. The pilA and pilB genes are in opposite orientation relative to each other, and the 5' ends of the two genes overlap. We discuss how these two loci may interact to control pilin expression in the gonococcus.
我们鉴定出了两个紧密连锁的基因,pilA和pilB,它们对菌毛蛋白启动子起反式作用。pilA - pilB定位于淋球菌染色体上表达位点pilE1和opaE1的下游。亚克隆数据表明,pilB对菌毛蛋白启动子起负向作用,pilB的插入失活会导致菌毛过多的淋球菌。一个pilA克隆可激活大肠杆菌中的菌毛蛋白启动子,并且一个pilA - /pilA +异源二倍体淋球菌表现出P - 表型。我们无法获得简单的pilA - 突变体,这强烈表明pilA是淋球菌中的一个必需基因。在体外偶联转录/翻译系统中,跨越pilA和pilB区域的插入片段指导合成两种分别为40和58 kd的蛋白质。DNA序列分析表明,pilA和pilB位点分别编码38.6 kd(具有一个假定的DNA结合结构域)和57.9 kd的蛋白质。pilA和pilB基因彼此方向相反,并且两个基因的5'端重叠。我们讨论了这两个位点如何相互作用以控制淋球菌中菌毛蛋白的表达。