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Conformational changes in deoxyribonucleic acid during transcription

W Wachsman, D D Anthony

    Biochemistry
    |December 23, 1980
    PubMed
    Summary

    Circular dichroism revealed DNA structural changes during transcription. RNA synthesis induced a B- to A-like DNA conformation shift, indicating altered secondary structure.

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    Area of Science:

    • Molecular Biology
    • Biophysics
    • Genetics

    Background:

    • DNA secondary structure is crucial for cellular processes.
    • In vitro transcription provides a model to study DNA-RNA polymerase interactions.
    • Circular dichroism (CD) is a sensitive technique for analyzing polynucleotide conformation.

    Purpose of the Study:

    • To investigate DNA secondary structure alterations during in vitro transcription.
    • To determine the role of RNA polymerase in DNA conformational changes.
    • To characterize the nature of CD spectral changes observed during RNA synthesis.

    Main Methods:

    • Circular dichroism (CD) spectroscopy was employed to monitor DNA secondary structure.
    • In vitro transcription assays were performed using calf thymus DNA and RNA polymerase.
    • Enzymatic treatments with RNase, restriction endonucleases, and DNase I were used to probe DNA structure.

    Main Results:

    • Formation of DNA-RNA polymerase complex did not affect DNA's CD spectrum.
    • RNA synthesis induced alterations in CD ellipticity (245-300 nm), suggesting a B- to A-like DNA conformational transition.
    • Two components of ellipticity change were identified: one DNA-intrinsic (component I) and one ribonuclease-labile (component II).
    • Component I was partially sensitive to DNase I, indicating DNA structural changes extending beyond polymerase binding sites.

    Conclusions:

    • In vitro transcription induces significant alterations in DNA secondary structure.
    • The observed B- to A-like transition is linked to RNA synthesis, not solely polymerase binding.
    • DNA structural changes during transcription are cooperative and extend beyond protected regions.

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