Espéli O, Moulin L, Boccard F
Centre de Génétique Moléculaire du CNRS, Centre National de la Recherche Scientifique, Gif-sur-Yvette Cedex, F-91198, France.
J Mol Biol. 2001 Nov 30;314(3):375-86. doi: 10.1006/jmbi.2001.5150.
Transcription attenuation comprises several processes that affect transcript elongation and transcription termination, and has an important role in regulating gene expression. In most cases, transcription attenuation is used as a regulatory mechanism that allows the cell to adjust protein synthesis levels in response to a specific signal. Here, by using a tRNA gene as a transcriptional reporter, we characterize a new type of transcription attenuation mechanism in Escherichia coli that involves bacterial interspersed mosaic elements (BIMEs), the main family of repetitive extragenic elements. The transcription termination factor Rho is required for attenuation in association with BIMEs, thus revealing a new role for Rho as a BIMEs-dependent global regulator of gene expression. By mutational analyses, we identified nucleotide determinants of BIMEs that are required for attenuation and showed that this process relies on a sequence-specific mechanism. Our data are consistent with a model in which BIMEs provoke a pause in RNA polymerase movement and Rho acts ultimately to terminate transcription. BIME-dependent transcription attenuation may be used as a means to differentially regulate expression of adjacent genes belonging to a single operon. BIMEs are dispersed in more than 250 operons such that attenuation can simultaneously affect expression of a large number of genes encoding unrelated proteins. This attenuation phenomenon, together with the known ability of BIMEs to stabilize upstream mRNA, reveals how dispersion of these abundant repetitive elements may affect gene regulation at the genome level.
转录衰减包含几个影响转录延伸和转录终止的过程,并且在调节基因表达中发挥重要作用。在大多数情况下,转录衰减用作一种调节机制,使细胞能够响应特定信号来调整蛋白质合成水平。在此,我们以一个tRNA基因为转录报告子,鉴定出大肠杆菌中一种新型的转录衰减机制,该机制涉及细菌散布镶嵌元件(BIMEs),即重复基因外元件的主要家族。转录终止因子Rho与BIMEs结合时对衰减是必需的,从而揭示了Rho作为一种依赖BIMEs的基因表达全局调节因子的新作用。通过突变分析,我们确定了衰减所需的BIMEs的核苷酸决定因素,并表明这一过程依赖于序列特异性机制。我们的数据与一个模型一致,即BIMEs引发RNA聚合酶移动的停顿,而Rho最终作用于终止转录。依赖BIMEs的转录衰减可能用作一种手段来差异调节属于单个操纵子的相邻基因的表达。BIMEs分散在250多个操纵子中,使得衰减能够同时影响大量编码不相关蛋白质的基因的表达。这种衰减现象,连同BIMEs稳定上游mRNA的已知能力,揭示了这些丰富的重复元件的分散如何可能在基因组水平上影响基因调控。