Parkinson J S, Houts S E
J Bacteriol. 1982 Jul;151(1):106-13. doi: 10.1128/jb.151.1.106-113.1982.
Six Escherichia coli che loci (cheA,-B,-R,-W,-Y, and Z) are located in two adjacent operons that map at minute 42 on the chromosome. Point mutants defective in any of these six functions have aberrant swimming patterns and are generally nonchemotactic. Deletions within the two major che gene operons were isolated in order to examine epistatic interactions among these genes. We first constructed a specialized transducing phage (lambda che22), which carries both of the che operons and their associated promoters. Deleted lambda che22 derivatives were selected by chelating agent inactivation, and these derivatives were characterized by mapping them against a series of host strains with point mutations. Representative nonpolar deletions were then transferred into the E. coli chromosome by homologous recombination. Although the phenotype of cheR mutants (smooth swimming) was expected to be epistatic to that of cheB mutants (tumbly swimming), we found that deletion mutants lacking both of these functions exhibited frequent directional changes or tumbling episodes as they swam. An examination of larger deletions indicated that both the cheA-cheW and cheY-cheZ functions were required for the anomalous tumbling behavior observed in these mutants. Loss of the cheB function was also correlated with an inverted behavioral response to sodium acetate, a strong repellent of wild-type cells. These findings indicate that an important component of the signal transducing machinery may be altered in cheB mutants.
六个大肠杆菌che基因座(cheA、-B、-R、-W、-Y和-Z)位于两个相邻的操纵子中,这些操纵子在染色体上的42分钟处定位。在这六种功能中任何一种有缺陷的点突变体都有异常的游动模式,并且通常是非趋化性的。为了研究这些基因之间的上位相互作用,分离了两个主要che基因操纵子内的缺失突变体。我们首先构建了一种特殊的转导噬菌体(λche22),它携带两个che操纵子及其相关启动子。通过螯合剂失活选择缺失的λche22衍生物,并通过将它们与一系列具有点突变的宿主菌株进行定位来表征这些衍生物。然后通过同源重组将代表性的非极性缺失转移到大肠杆菌染色体中。尽管预期cheR突变体(平滑游动)的表型对cheB突变体(翻滚游动)的表型是上位的,但我们发现缺乏这两种功能的缺失突变体在游动时表现出频繁的方向变化或翻滚事件。对更大缺失的研究表明,cheA-cheW和cheY-cheZ功能对于在这些突变体中观察到的异常翻滚行为都是必需的。cheB功能的丧失也与对乙酸钠(野生型细胞的强驱避剂)的反向行为反应相关。这些发现表明,信号转导机制的一个重要组成部分可能在cheB突变体中发生了改变。