Loewy Z G, Bryan R A, Reuter S H, Shapiro L
Albert Einstein College of Medicine, Department of Molecular Biology, Bronx, New York 10461.
Genes Dev. 1987 Aug;1(6):626-35. doi: 10.1101/gad.1.6.626.
The Caulobacter crescentus flagellum is assembled during a defined time period in the cell cycle. Two genes encoding the major components of the flagellar filament, the 25K and the 27.5K flagellins, are expressed coincident with flagellar assembly. A third gene, flgJ, is also temporally regulated. The synthesis of the product of flgJ, the 29K flagellin, occurs prior to the synthesis of the other flagellin proteins. We demonstrate here that the time of initiation of flgJ expression is independent of chromosomal location but is dependent upon cis-acting sequences present upstream of the flgJ structural gene. Evidence that there is transcriptional control of flgJ expression includes the following: (1) The initial appearance of flgJ message was coincident with the onset of 29K flagellin protein synthesis, and (2) expression of an NPT II reporter gene driven by the flgJ promoter was temporally correct. Post-transcriptional regulation might contribute to the control of expression, because the flgJ mRNA persisted for a longer period of time than did the synthesis of the 29K protein. The 29K flagellin was found only in the progeny swarmer cell after cell division. In a mutant strain that failed to assemble a flagellum, the 29K flagellin still segregated to the presumptive swarmer cell, demonstrating that positioning of the protein is independent of filament assembly. Analysis of a chimeric flgJ-NPT II transcriptional fusion showed that the flgJ regulatory sequences do not control the segregation of the 29K flagellin to the swarmer cell progeny, suggesting that correct segregation depends on the protein product.
新月柄杆菌鞭毛在细胞周期的特定时间段内组装。两个编码鞭毛丝主要成分的基因,即25K和27.5K鞭毛蛋白,在鞭毛组装时同时表达。第三个基因flgJ也受到时间调控。flgJ产物29K鞭毛蛋白的合成发生在其他鞭毛蛋白合成之前。我们在此证明,flgJ表达的起始时间与染色体位置无关,但依赖于flgJ结构基因上游存在的顺式作用序列。flgJ表达存在转录控制的证据如下:(1)flgJ信息的最初出现与29K鞭毛蛋白合成的开始同时发生,(2)由flgJ启动子驱动的NPT II报告基因的表达在时间上是正确的。转录后调控可能有助于表达控制,因为flgJ mRNA持续的时间比29K蛋白的合成时间更长。29K鞭毛蛋白仅在细胞分裂后的子代游动细胞中发现。在一个无法组装鞭毛的突变菌株中,29K鞭毛蛋白仍会分离到假定的游动细胞中,这表明该蛋白的定位与鞭毛丝组装无关。对嵌合flgJ-NPT II转录融合体的分析表明,flgJ调控序列并不控制29K鞭毛蛋白向游动细胞子代的分离,这表明正确的分离取决于蛋白产物。