Aberg Anna, Shingler Victoria, Balsalobre Carlos
Department of Molecular Biology, Umeå University, S-90187 Umeå, Sweden.
Mol Microbiol. 2008 Mar;67(6):1223-41. doi: 10.1111/j.1365-2958.2008.06115.x. Epub 2008 Feb 13.
The expression of type 1 fimbriae is dependent on the intracellular levels of ppGpp through stimulation of fimB transcription. Here we show that in contrast to the previously described decreased fimbriation observed in a ppGpp-deficient strain, DksA deficiency results in a hyperfimbriated state. In vivo assays show that the effect of DksA deficiency on the type 1 fimbriae occurs at the phase variation level because of elevated transcription from the fimB P2 promoter. In contrast, our in vitro transcription studies demonstrate that ppGpp and DksA can stimulate transcription from the fimB P2 promoter both independently and codependently. We provide evidences that the apparently contradictory results from the in vivo and in vitro transcriptional studies are at least in part a consequence of the increased association of the anti-pausing factors (GreA and GreB) to the RNA polymerase in the absence of DksA in vivo.
1型菌毛的表达通过刺激fimB转录而依赖于ppGpp的细胞内水平。在这里我们表明,与之前描述的在ppGpp缺陷菌株中观察到的菌毛形成减少相反,DksA缺陷导致菌毛过度形成状态。体内试验表明,DksA缺陷对1型菌毛的影响发生在相变水平,这是由于fimB P2启动子转录增加所致。相比之下,我们的体外转录研究表明,ppGpp和DksA可以独立地和相互依赖地刺激fimB P2启动子的转录。我们提供的证据表明,体内和体外转录研究中明显矛盾的结果至少部分是由于体内缺乏DksA时抗暂停因子(GreA和GreB)与RNA聚合酶的结合增加所致。