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RNA polymerase binding sites in lambdaplac5 DNA.

B B Jones, H Chan, S Rothstein

    Proceedings of the National Academy of Sciences of the United States of America
    |November 1, 1977
    PubMed
    Summary
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    Escherichia coli RNA polymerase binds to specific promoter sites on lambdaplac5 DNA. Binding also occurs in non-promoter regions, potentially involving termination sites and A-T rich DNA sequences.

    Area of Science:

    • Molecular Biology
    • Genetics
    • Biochemistry

    Background:

    • Escherichia coli RNA polymerase is crucial for gene transcription.
    • Understanding DNA-RNA polymerase interactions is key to gene regulation.

    Purpose of the Study:

    • To investigate the in vitro binding sites of E. coli RNA polymerase on lambdaplac5 DNA fragments.
    • To identify specific DNA sequences and regions recognized by RNA polymerase.

    Main Methods:

    • Filter binding technique using restriction endonuclease-generated DNA fragments (HindII, HindIII).
    • Analysis of DNA cleavage patterns after RNA polymerase binding to map additional sites.

    Main Results:

    • RNA polymerase binding confirmed at known promoters (p(R)', p(I), mis, p(O), p(rm)).

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  • Binding also observed in non-promoter regions (b2, cIII-t(L)) and potential termination sites.
  • Additional binding sites identified using DNA cleavage inhibition, including p(L) and p(R).
  • Binding sites correlate with A-T rich regions in the lambda genome.

    • Conclusions:

    • RNA polymerase binding is influenced by specific recognition sequences.
    • DNA helix stability in A-T rich regions also plays a role in polymerase binding.
    • Identified novel binding sites beyond canonical promoters.