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Related Experiment Videos

Palindromic sequences preceding the terminator increase polymerase III template activity

W M Chu1, R E Ballard, C W Schmid

  • 1Section of Molecular and Cellular Biology, University of California, Davis, CA 95616, USA.

Nucleic Acids Research
|June 1, 1997
PubMed
Summary

A palindromic sequence preceding a terminator enhances RNA polymerase III (pol III) transcription. Mutations disrupting this sequence

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Area of Science:

  • Molecular Biology
  • Gene Regulation
  • Biochemistry

Background:

  • RNA polymerase III (pol III) transcribes genes encoding small RNAs.
  • Pol III transcription termination typically involves a sequence of four thymidine (T) residues.
  • Previous studies indicated that a palindromic sequence upstream of the T-rich terminator enhances transcription.

Purpose of the Study:

  • To investigate the role of the palindromic sequence's dyad symmetry in RNA polymerase III (pol III) transcription termination.
  • To determine if the higher-order structure preceding the terminator influences pol III activity.

Main Methods:

  • Site-directed mutagenesis to alter the dyad symmetry of the palindromic sequence.
  • In vitro and in vivo assays to measure transcriptional expression.

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  • Compensatory mutations to restore dyad symmetry and assess rescue of transcriptional activity.

    • Main Results:

    • Mutations eliminating dyad symmetry in the palindromic sequence decreased transcriptional expression.
    • This effect was observed regardless of whether symmetry was disrupted proximally or distally to the terminator.
    • Restoring dyad symmetry through compensatory mutations rescued transcriptional activity.

    Conclusions:

    • The dyad symmetry of the palindromic sequence is crucial for enhancing pol III transcriptional activity.
    • A higher-order RNA structure, likely a hairpin, formed by the palindromic sequence immediately upstream of the terminator increases pol III transcription.