期刊
JOURNAL OF BIOLOGICAL CHEMISTRY
卷 286, 期 25, 页码 22600-22608出版社
AMER SOC BIOCHEMISTRY MOLECULAR BIOLOGY INC
DOI: 10.1074/jbc.M111.247080
关键词
-
资金
- National Institutes of Health [R01 GM64530]
- Russian Academy of Sciences Presidium
- Federal Program Scientific and Scientific-Pedagogical Personnel of Innovative Russia [02.740.11.0771]
Nucleation of promoter melting in bacteria is coupled with RNA polymerase (RNAP) binding to a conserved - 10 promoter element located at the upstream edge of the transcription bubble. The mechanism of downstream propagation of the transcription bubble to include the transcription start site is unclear. Here we introduce new model downstream fork junction promoter fragments that specifically bind RNAP and mimic the downstream segment of promoter complexes. We demonstrate that RNAP binding to downstream fork junctions is coupled with DNA melting around the transcription start point. Consequently, certain downstream fork junction probes can serve as transcription templates. Using a protein beacon fluorescent method, we identify structural determinants of affinity and transcription activity of RNAP-downstream fork junction complexes. Measurements of RNAP interaction with double-stranded promoter fragments reveal that the strength of RNAP interactions with downstream DNA plays a critical role in promoter opening and that the length of the downstream duplex must exceed a critical length for efficient formation of transcription competent open promoter complex.
作者
我是这篇论文的作者
点击您的名字以认领此论文并将其添加到您的个人资料中。
推荐
暂无数据