Mueller K, Oebbecke C, Förster G
Cell. 1977 Jan;10(1):121-30. doi: 10.1016/0092-8674(77)90146-5.
The rate of in vitro transcription of the rRNA genes of E. coli is more than 20 fold higher than the averaged transcription rate of other genome segments of the same size. This "preferential transcription" of rRNA genes reflects a high efficiency of their promoters in chain initiation. We show that the high initiation rate at rRNA promoters results from a high rate of RNA polymerase binding to these promoters as measured by the formation of heparin-resistant RNA polymerase-DNA complexes. The results indicate that the preferential binding of RNA polymerase to rRNA promoters is mainly due to their large binding capacity rather than to a high rate constant of polymerase binding to a single binding site. The polymerase binding capcity of rRNA promoters was estimated from the number of rRNA chains initiated by heparin-resistant complexes under conditions of template saturation and from the number of rRNA transcription units participating in the binding reaction. At least 30 RNA polymerase molecules were found to be protected from heparin per rRNA transcription unit. The rest of the genome (99.4%; possibly sufficient to encode 4000 nonribosomal RNA species) protects under these conditions 2000 enzyme molecules. These results suggest that a high multiplicity of RNA polymerase binding may be responsible for the high efficiency of rRNA promoters. The validity of this hypothesis is discussed.
大肠杆菌rRNA基因的体外转录速率比相同大小的其他基因组片段的平均转录速率高20多倍。rRNA基因的这种“优先转录”反映了其启动子在链起始方面的高效率。我们发现,通过形成对肝素耐药的RNA聚合酶-DNA复合物来测量,rRNA启动子处的高起始速率源于RNA聚合酶与这些启动子的高结合速率。结果表明,RNA聚合酶与rRNA启动子的优先结合主要是由于它们较大的结合能力,而不是由于聚合酶与单个结合位点的高结合速率常数。rRNA启动子的聚合酶结合能力是根据在模板饱和条件下由对肝素耐药的复合物起始的rRNA链的数量以及参与结合反应的rRNA转录单位的数量来估计的。发现每个rRNA转录单位至少有30个RNA聚合酶分子受到肝素的保护。基因组的其余部分(99.4%;可能足以编码4000种非核糖体RNA种类)在这些条件下保护2000个酶分子。这些结果表明,RNA聚合酶结合的高多重性可能是rRNA启动子高效率的原因。讨论了这一假设的有效性。