Open AccessMinus-strand origin of filamentous phage versus transcriptional promoters in recognition of RNA polymerase
Author(s) -
Akihiro Higashitani,
Nahoko Higashitani,
Kensuke Horiuchi
Publication year - 1997
Publication title -
proceedings of the national academy of sciences of the united states of america
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 5.011
H-Index - 771
eISSN - 1091-6490
pISSN - 0027-8424
DOI - 10.1073/pnas.94.7.2909
Subject(s) - rna polymerase , microbiology and biotechnology , coding strand , polymerase , transcription (linguistics) , biology , rna dependent rna polymerase , rna , promoter , primer (cosmetics) , dna , transcription bubble , rna polymerase i , rna polymerase ii , specificity factor , biochemistry , chemistry , gene , gene expression , linguistics , philosophy , organic chemistry
Replication of complementary-strand DNA in filamentous phages is initiated by a primer RNA that is synthesized at the minus-strand origin on the viral single-stranded DNA byEscherichia coli RNA polymerase holoenzyme containing the σ70 subunit. We have demonstrated that the affinity of RNA polymerasein vitro to the origin is about 16-fold higher than that to thelac UV5 promoter. We have also shown that the temperature dependence of the primer RNA synthesis is much lower than that oflac UV5 transcription. The high affinity of RNA polymerase to the origin depends on the single strandedness of the “−10 region.” A nucleotide sequence of the nontemplate strand in the −10 region was found to be important for the function, but that of the template strand was not. These observations suggest that σ70 subunit directly interacts with the single-stranded nontemplate strand containing adenine residue(s) at the −10 region of promoter.