Evidence for a regulatory function of the histone-like Escherichia coli protein H-NS in ribosomal RNA synthesis.
We have isolated a small Escherichia coli protein which stably interacts with ribosomal RNA P1 promoter DNA. We present evidence showing that the protein is identical to the histone-like E. coli protein, H-NS (H1). Binding of H-NS to the P1 promoter region is dependent on the DNA curvature. Mapping the H-NS-DNA contact sites by nuclease protection and high-resolution footprinting techniques reveal three H-NS-binding domains, and contacts of the protein in the major groove of the bent DNA. The binding region extends from position -18 to -89, relative to the P1 transcription start site, and shows an overlap with the known binding sites for Fis, another E. coli protein, which acts as transcriptional activator of P1. The binding of H-NS does not displace Fis; instead, heterologous complexes are formed. Apparently, H-NS and Fis bind to separated curved DNA segments, with the planes of the curves pointing into different directions. In vitro transcriptional analyses demonstrate that H-NS represses rRNA P1 promoter-directed transcription. Repression is most pronounced in the presence of Fis. Thus, H-NS seems specifically to antagonize Fis-dependent activation. No comparable inactivation is observed for the second rRNA promoter P2.
Tippner
D
D
Institut für Physikalische Biologie, Heinrich-Heine-Universität Düsseldorf, Germany.
Afflerbach
H
H
Bradaczek
C
C
Wagner
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R
D001425:Bacterial Outer Membrane Proteins / Q000378:metabolism*; D001426:Bacterial Proteins; D001483:Base Sequence; D001667:Binding, Competitive; D004269:DNA, Bacterial / Q000737:chemistry / Q000378:metabolism; D004268:DNA-Binding Proteins / Q000378:metabolism*; D004926:Escherichia coli / Q000235:genetics* / Q000378:metabolism; D015964:Gene Expression Regulation, Bacterial; D008969:Molecular Sequence Data; D009690:Nucleic Acid Conformation; D011401:Promoter Regions, Genetic; D012329:RNA, Bacterial / Q000096:biosynthesis*; D012335:RNA, Ribosomal / Q000096:biosynthesis*; D014158:Transcription, Genetic
D001425:Bacterial Outer Membrane Proteins; D001426:Bacterial Proteins; D004269:DNA, Bacterial; D004268:DNA-Binding Proteins; C031908:H-NS protein, bacteria; D012329:RNA, Bacterial; D012335:RNA, Ribosomal