Wingrove J A, Gober J W
Department of Chemistry and Biochemistry, University of California, Los Angeles 90024.
Genes Dev. 1994 Aug 1;8(15):1839-52. doi: 10.1101/gad.8.15.1839.
The differential localization of proteins in the Caulobacter predivisional cell leads to the formation of two distinct progeny cells: a motile swarmer cell and a sessile stalked cell. Pole-specific transcription in the predivisional cell is one mechanism responsible for protein localization. Here we show that the sigma 54 transcriptional activator FlbD, which activates swarmer pole-specific transcription of a subset of late flagellar genes, is also capable of functioning as a pole-specific repressor of the early flagellar fliF operon. DNase I footprinting and methylation interference assays indicate that FlbD binds to regions of the fliF promoter at regions that would be likely to interfere with the binding of RNA polymerase. A mutation that abolishes FlbD binding results in up to a fourfold increase in fliF promoter expression. This mutation alters both the spatial and temporal pattern of fliF expression resulting in the inappropriate expression of the fliF operon in the swarmer pole of the predivisional cell. These results demonstrate that FlbD represses early flagellar gene expression in the swarmer pole of the Caulobacter predivisional cell. This is the first instance in which a protein specifically involved in pole-specific repression has been identified in Caulobacter. The restriction of FlbD activity to the swarmer pole accomplishes two regulatory missions by simultaneously activating late flagellar gene expression and repressing early flagellar genes.
一个游动的游动细胞和一个固着的柄细胞。前分裂细胞中的极特异性转录是负责蛋白质定位的一种机制。在这里,我们表明,激活鞭毛晚期基因子集的游动极特异性转录的σ54转录激活因子FlbD,也能够作为早期鞭毛fliF操纵子的极特异性阻遏物发挥作用。DNA酶I足迹法和甲基化干扰试验表明,FlbD结合到fliF启动子的区域,这些区域可能会干扰RNA聚合酶的结合。消除FlbD结合的突变导致fliF启动子表达增加高达四倍。该突变改变了fliF表达的空间和时间模式,导致fliF操纵子在前分裂细胞的游动极中不适当表达。这些结果表明,FlbD抑制柄杆菌属前分裂细胞游动极中的早期鞭毛基因表达。这是首次在柄杆菌属中鉴定出一种特异性参与极特异性阻遏的蛋白质。将FlbD活性限制在游动极通过同时激活晚期鞭毛基因表达和抑制早期鞭毛基因完成了两个调控任务。