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Direct Restart of a Replication Fork Stalled by a Head-On RNA Polymerase
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DNA replication fork pause sites dependent on transcription

A M Deshpande1, C S Newlon

  • 1Department of Micobiology and Molecular Genetics, UMDNJ Medical School and UMDNJ-Graduate School of Biomedical Sciences, Newark, NJ 07103, USA.

Science (New York, N.Y.)
|May 17, 1996
PubMed

Insights

Replication fork pause sites in yeast were found to be located in transfer RNA genes. These sites stall DNA replication forks only when opposing transcription, indicating that transcription is essential for this process.

Area of Science:

  • Molecular Biology
  • Genetics
  • Biochemistry

Background:

  • Replication fork movement can be transiently arrested at specific DNA sites.
  • Transfer RNA (tRNA) genes are known to play roles in genome stability.

Purpose of the Study:

  • To identify the genomic locations of replication fork pause (RFP) sites in Saccharomyces cerevisiae.
  • To investigate the relationship between tRNA gene transcription and replication fork stalling.

Main Methods:

  • In vivo mapping of replication fork pause sites.
  • Analysis of mutant tRNA genes with defects in transcription initiation.
  • Utilizing a temperature-sensitive RNA polymerase III mutant (rpc160-41).

Main Results:

  • Replication fork pause sites were localized to transfer RNA (tRNA) genes.
  • RFP sites exhibit polarity, stalling forks only when they oppose the direction of tRNA transcription.
  • Mutant tRNA genes and RNA polymerase III mutants defective in transcription initiation did not cause replication fork stalling.

Conclusions:

  • Transcription of tRNA genes is required for the activity of replication fork pause sites.
  • The findings suggest a direct link between the transcription machinery and the regulation of DNA replication fork progression at tRNA genes.

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