Gober J W, Shapiro L
Department of Developmental Biology, Beckman Center, Stanford University School of Medicine, California 94305-5427.
Mol Biol Cell. 1992 Aug;3(8):913-26. doi: 10.1091/mbc.3.8.913.
The transcription of a group of flagellar genes is temporally and spatially regulated during the Caulobacter crescentus cell cycle. These genes all share the same 5' cis-regulatory elements: a sigma 54 promoter, a binding site for integration host factor (IHF), and an enhancer sequence, known as the ftr element. We have partially purified the ftr-binding proteins, and we show that they require the same enhancer sequences for binding as are required for transcriptional activation. We have also partially purified the Caulobacter homolog of IHF and demonstrate that it can facilitate in vitro integrase-mediated lambda recombination. Using site-directed mutagenesis, we provide the first demonstration that natural enhancer sequences and IHF binding elements that reside 3' to the sigma 54 promoter of a bacterial gene, flaNQ, are required for transcription of the operon, in vivo. The IHF protein and the ftr-binding protein is primarily restricted to the predivisional cell, the cell type in which these promoters are transcribed. flaNQ promoter expression is localized to the swarmer pole of the predivisional cell, as are other flagellar promoters that possess these regulatory sequences 5' to the start site. The requirement for an IHF binding site and an ftr-enhancer element in spatially transcribed flagellar promoters indicates that a common mechanism may be responsible for both temporal and polar transcription.
在新月柄杆菌的细胞周期中,一组鞭毛基因的转录在时间和空间上受到调控。这些基因都共享相同的5'顺式调控元件:一个σ⁵⁴启动子、一个整合宿主因子(IHF)的结合位点以及一个增强子序列,即ftr元件。我们已经部分纯化了ftr结合蛋白,并且我们表明它们结合所需的增强子序列与转录激活所需的序列相同。我们还部分纯化了IHF的柄杆菌同源物,并证明它可以促进体外整合酶介导的λ重组。使用定点诱变,我们首次证明,细菌基因flaNQ的σ⁵⁴启动子3'端的天然增强子序列和IHF结合元件在体内对于操纵子的转录是必需的。IHF蛋白和ftr结合蛋白主要局限于分裂前细胞,即转录这些启动子的细胞类型。flaNQ启动子的表达定位于分裂前细胞的游动极,其他具有这些位于起始位点5'端调控序列的鞭毛启动子也是如此。在空间转录的鞭毛启动子中对IHF结合位点和ftr增强子元件的需求表明,一种共同机制可能负责时间和极性转录。