Ichida Hiroyuki, Long Sharon R
Department of Biology, Stanford University, Stanford, CA 94305, USA RIKEN Nishina Center for Accelerator-Based Science, Wako, Saitama 351-0198, Japan.
Department of Biology, Stanford University, Stanford, CA 94305, USA
Nucleic Acids Res. 2016 Jun 20;44(11):5045-53. doi: 10.1093/nar/gkw435. Epub 2016 May 17.
Identifying functional elements in promoter sequences is a major goal in computational and experimental genome biology. Here, we describe an algorithm, Local Distribution of Short Sequences for Prokaryotes (LDSS-P), to identify conserved short motifs located at specific positions in the promoters of co-expressed prokaryotic genes. As a test case, we applied this algorithm to a symbiotic nitrogen-fixing bacterium, Sinorhizobium meliloti The LDSS-P profiles that overlap with the 5' section of the extracytoplasmic function RNA polymerase sigma factor RpoE2 consensus sequences displayed a sharp peak between -34 and -32 from TSS positions. The corresponding genes overlap significantly with RpoE2 targets identified from previous experiments. We further identified several groups of genes that are co-regulated with characterized marker genes. Our data indicate that in S. meliloti, and possibly in other Rhizobiaceae species, the master cell cycle regulator CtrA may recognize an expanded motif (AACCAT), which is positionally shifted from the previously reported CtrA consensus sequence in Caulobacter crescentus Bacterial one-hybrid experiments showed that base substitution in the expanded motif either increase or decrease the binding by CtrA. These results show the effectiveness of LDSS-P as a method to delineate functional promoter elements.
识别启动子序列中的功能元件是计算和实验基因组生物学的一个主要目标。在此,我们描述了一种算法,即原核生物短序列局部分布算法(LDSS-P),用于识别共表达原核基因启动子中特定位置的保守短基序。作为一个测试案例,我们将该算法应用于共生固氮细菌苜蓿中华根瘤菌。与胞外功能RNA聚合酶西格玛因子RpoE2共有序列5' 部分重叠的LDSS-P图谱在转录起始位点(TSS)位置的 -34至 -32之间显示出一个尖峰。相应的基因与先前实验中鉴定的RpoE2靶标有显著重叠。我们进一步鉴定了几组与特征性标记基因共同调控的基因。我们的数据表明,在苜蓿中华根瘤菌以及可能在其他根瘤菌科物种中,主细胞周期调节因子CtrA可能识别一个扩展基序(AACCAT),该基序在位置上与先前报道的新月柄杆菌中的CtrA共有序列有所偏移。细菌单杂交实验表明,扩展基序中的碱基替换要么增加要么减少CtrA的结合。这些结果表明LDSS-P作为一种描绘功能性启动子元件的方法是有效的。