Albrechtsen B, Ross B M, Squires C, Squires C L
Department of Biological Sciences, Columbia University, New York, NY 10027.
Nucleic Acids Res. 1991 Apr 25;19(8):1845-52. doi: 10.1093/nar/19.8.1845.
We have examined the termination region sequence of the rrnG operon and have observed its properties in vivo using a fusion plasmid test system. Transcription of rrnG terminator fragments was also studied in vitro. We found that termination of rrnG transcription is a complex process controlled by a tandem Rho-independent and Rho-dependent terminator arrangement which we designate rrnG-tt'. Together, these two elements were 98% efficient at terminating transcription initiated at the rrnG-P2 promoter. When the two elements were separated, however, we found that the Rho-independent structure was only 59% efficient while the Rho-dependent fragment alone could account for total transcriptional termination of the tandem arrangement. The rrnG termination region was resistant to rrn antitermination and, therefore, possesses some means of stopping antiterminated transcription. The distal rrnG sequence contains several additional noteworthy features; the rrnGt' fragment contains a REP (repetitive extragenic palindromic) sequence and homology with a small unidentified reading frame following rrnE. This sequence is followed by witA, which is homologous to a citrate transport gene, citB. Finally, our sequence, obtained from plasmid pLC23-30, contains a Tn1000 insertion that is absent from the E. coli chromosome. This insertion lies 975 bp beyond the 5S gene and is not involved in the termination events examined in this study.
我们研究了rrnG操纵子的终止区域序列,并使用融合质粒测试系统在体内观察了其特性。还在体外研究了rrnG终止子片段的转录。我们发现rrnG转录的终止是一个复杂的过程,由串联的不依赖Rho和依赖Rho的终止子排列控制,我们将其命名为rrnG-tt'。这两个元件共同作用时,在终止从rrnG-P2启动子起始的转录方面效率为98%。然而,当这两个元件分开时,我们发现不依赖Rho的结构效率仅为59%,而单独的依赖Rho的片段就能导致串联排列的完全转录终止。rrnG终止区域对rrn抗终止具有抗性,因此具有某种阻止抗终止转录的手段。rrnG的远端序列包含几个其他值得注意的特征;rrnGt'片段包含一个REP(重复基因外回文序列)序列,并且与rrnE之后一个未鉴定的小阅读框具有同源性。该序列之后是witA,它与一个柠檬酸盐转运基因citB同源。最后,我们从质粒pLC23-30获得的序列包含一个大肠杆菌染色体中不存在的Tn1000插入。这个插入位于5S基因下游975 bp处,并且不参与本研究中检测的终止事件。