Nodwell J R, Greenblatt J
Banting and Best Department of Medical Research, University of Toronto, Canada.
Genes Dev. 1991 Nov;5(11):2141-51. doi: 10.1101/gad.5.11.2141.
The boxA and boxB components of the lambda nut site are important for transcriptional antitermination by the phage N protein. We show here that boxA and boxB RNA in N-modified transcription complexes are inaccessible to ribonucleases and have altered sensitivity to dimethylsulfate. N and NusA suffice to weakly protect boxB, independently of boxA and other factors. However, efficient protection of the entire nut site from ribonucleases requires boxA and boxB, N, NusA, NusB, S10, and NusG. Mutations in RNA polymerase, which inhibit antitermination by N in vivo, disallow protection of the nut site during transcription in vitro; therefore, the surface of RNA polymerase must coordinate the formation of complexes containing the antitermination factors and nut site RNA.
λ噬菌体nut位点的boxA和boxB组件对于噬菌体N蛋白介导的转录抗终止作用很重要。我们在此表明,在N修饰的转录复合物中的boxA和boxB RNA对核糖核酸酶不可接近,并且对硫酸二甲酯的敏感性发生了改变。N和NusA足以对boxB提供弱保护,这与boxA和其他因素无关。然而,要有效保护整个nut位点免受核糖核酸酶的作用,则需要boxA和boxB、N、NusA、NusB、S10和NusG。RNA聚合酶中的突变在体内会抑制N介导的抗终止作用,在体外转录过程中也不允许对nut位点进行保护;因此,RNA聚合酶的表面必须协调包含抗终止因子和nut位点RNA的复合物的形成。