Mooney Rachel A, Davis Sarah E, Peters Jason M, Rowland Jennifer L, Ansari Aseem Z, Landick Robert
Department of Biochemistry, University of Wisconsin, Madison, WI 53706, USA.
Mol Cell. 2009 Jan 16;33(1):97-108. doi: 10.1016/j.molcel.2008.12.021.
The trafficking patterns of the bacterial regulators of transcript elongation sigma(70), rho, NusA, and NusG on genes in vivo and the explanation for promoter-proximal peaks of RNA polymerase (RNAP) are unknown. Genome-wide, E. coli ChIP-chip revealed distinct association patterns of regulators as RNAP transcribes away from promoters (rho first, then NusA, then NusG). However, the interactions of elongating complexes with these regulators did not differ significantly among most transcription units. A modest variation of NusG signal among genes reflected increased NusG interaction as transcription progresses, rather than functional specialization of elongating complexes. Promoter-proximal RNAP peaks were offset from sigma(70) peaks in the direction of transcription and co-occurred with NusA and rho peaks, suggesting that the RNAP peaks reflected elongating, rather than initiating, complexes. However, inhibition of rho did not increase RNAP levels within genes downstream from the RNAP peaks, suggesting the peaks are caused by a mechanism other than rho-dependent attenuation.
细菌转录延伸调节因子sigma(70)、rho、NusA和NusG在体内基因上的分布模式以及RNA聚合酶(RNAP)启动子近端峰的成因尚不清楚。全基因组范围的大肠杆菌染色质免疫沉淀芯片(ChIP-chip)显示,随着RNAP转录离开启动子,调节因子呈现出不同的关联模式(rho最先出现,然后是NusA,接着是NusG)。然而,在大多数转录单元中,延伸复合物与这些调节因子的相互作用并无显著差异。基因间NusG信号的适度变化反映出随着转录进行,NusG相互作用增强,而非延伸复合物的功能特化。启动子近端的RNAP峰在转录方向上与sigma(70)峰偏移,并与NusA和rho峰同时出现,这表明RNAP峰反映的是延伸复合物,而非起始复合物。然而,抑制rho并没有增加RNAP峰下游基因内的RNAP水平,这表明这些峰是由rho依赖性衰减以外的机制引起的。