THE INFLUENCE OF AN ALTERNATE TEMPLATE CONFORMATION ON ELONGATING PHAGE T7-RNA POLYMERASE

被引:15
作者
DROGE, P [1 ]
POHL, FM [1 ]
机构
[1] UNIV CONSTANCE,DEPT BIOL,W-7750 CONSTANCE,GERMANY
关键词
D O I
10.1093/nar/19.19.5301
中图分类号
Q5 [生物化学]; Q7 [分子生物学];
学科分类号
071010 ; 081704 ;
摘要
We investigated the effect of left-handed Z-DNA on transcription by bacteriophage T7 RNA polymerase in vitro and, surprisingly, found that the enzyme can efficiently utilize a template containing a stretch of left-handed DNA close to the promoter. Analysis of transcription products revealed that only a small fraction of elongating polymerases abort transcription either at the promoter proximal or at the distal B-to-Z junction and, even less frequently, within the stretch of left-handed DNA. Our results indicate that, unlike Ecoli RNA polymerase (ref. 1,2), T7 RNA polymerase can utilize a template with a CG stretch in an alternate conformation. In contrast, polymerases are completely blocked at the promoter proximal junction by a monoclonal antibody directed against Z-DNA. This blockage remains stable over a remarkable time, even when negative supercoiling is released by linearization of the template. Together with our recent finding of transcription-induced formation of Z-DNA (3), our data provide an example for a possible auto-regulatory mechanism that employs a change in DNA conformation.
引用
收藏
页码:5301 / 5306
页数:6
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