Department of Molecular Genetics, Weizmann Institute of Science, Rehovot, Israel.
PLoS One. 2013 Apr 16;8(4):e61850. doi: 10.1371/journal.pone.0061850. Print 2013.
Bdellovibrio bacteriovorus is an obligate predator of bacteria ubiquitously found in the environment. Its life cycle is composed of two essential phases: a free-living, non-replicative, fast swimming attack phase (AP) wherein the predator searches for prey; and a non-motile, actively dividing growth phase (GP) in which it consumes the prey. The molecular regulatory mechanisms governing the switch between AP and GP are largely unknown. We used RNA-seq to generate a single-base-resolution map of the Bdellovibrio transcriptome in AP and GP, revealing a specific "AP" transcriptional program, which is largely mutually exclusive of the GP program. Based on the expression map, most genes in the Bdellovibrio genome are classified as "AP only" or "GP only". We experimentally generated a genome-wide map of 140 AP promoters, controlling the majority of AP-specific genes. This revealed a common sigma-like DNA binding site highly similar to the E. coli flagellar genes regulator sigma28 (FliA). Further analyses suggest that FliA has evolved to become a global AP regulator in Bdellovibrio. Our results also reveal a non-coding RNA that is massively expressed in AP. This ncRNA contains a c-di-GMP riboswitch. We suggest it functions as an intracellular reservoir for c-di-GMP, playing a role in the rapid switch from AP to GP.
蛭弧菌是一种广泛存在于环境中的细菌的专性捕食者。它的生命周期由两个必要的阶段组成:一个自由生活的、非复制的、快速游动的攻击阶段(AP),在这个阶段,捕食者寻找猎物;和一个非运动的、主动分裂的生长阶段(GP),在这个阶段它消耗猎物。控制 AP 和 GP 之间转换的分子调节机制在很大程度上是未知的。我们使用 RNA-seq 生成了 Bdellovibrio 在 AP 和 GP 中的转录组的单碱基分辨率图谱,揭示了一个特定的“AP”转录程序,该程序在很大程度上与 GP 程序是相互排斥的。基于表达图谱,Bdellovibrio 基因组中的大多数基因被分类为“AP 仅”或“GP 仅”。我们实验生成了一个包含 140 个 AP 启动子的全基因组图谱,这些启动子控制着大多数 AP 特异性基因。这揭示了一个高度类似于大肠杆菌鞭毛基因调控因子 sigma28(FliA)的常见 sigma 样 DNA 结合位点。进一步的分析表明,FliA 已经进化成为 Bdellovibrio 中的一个全局 AP 调节剂。我们的研究结果还揭示了一种在 AP 中大量表达的非编码 RNA。这种 ncRNA 含有 c-di-GMP 核糖开关。我们认为它作为 c-di-GMP 的细胞内储存库发挥作用,在从 AP 到 GP 的快速转换中发挥作用。