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Journal ArticleDOI

Need for DNA topoisomerase activity as a swivel for DNA replication for transcription of ribosomal RNA.

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TLDR
It is reported that DNA and ribosomal RNA synthesis are drastically inhibited in an S. cerevisiae top1 top2 ts double mutant at the restrictive temperature, but that the rate of poly(A)+ RNA synthesis is reduced only about threefold and transfer RNA synthesis remains relatively normal.
Abstract
Yeast strains with mutations in the genes for DNA topoisomerases I and II have been identified previously in both Saccharomyces cerevisiae1–3 and Schizosaccharomyces pombe4. The topoisomerase II mutants (top2) are conditional-lethal temperature-sensitive (ts) mutants. They are defective in the termination of DNA replication and the segregation of daughter chromosomes2–5, but otherwise appear to replicate and transcribe DNA normally. Topoisomerase I mutants (top1), including strains with null mutations are viable and exhibit no obvious growth defects, demonstrating that DNA topoisomerase I is not essential for viability in yeast1,4,6,7. In contrast to the single mutants, top1 top2 ts double mutants from both Schizosaccharomyces pombe and Saccharomyces cerevisiae grow poorly at the permissive temperature and stop growth rapidly at the non-permissive temperature4,7. Here we report that DNA and ribosomal RNA synthesis are drastically inhibited in an S. cerevisiae top1 top2 ts double mutant at the restrictive temperature, but that the rate of poly(A)+ RNA synthesis is reduced only about threefold and transfer RNA synthesis remains relatively normal. The results suggest that DNA replication and at least ribosomal RNA synthesis require an active topoisomerase, presumably to act as a swivel to relieve torsional stress, and that either topoisomerase can perform the required function (except in termination of DNA replication where topoisomerase II is required).

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Journal ArticleDOI

Multiple Pathways of Recombination Induced by Double-Strand Breaks in Saccharomyces cerevisiae

TL;DR: This review encompasses different aspects of DSB-induced recombination in Saccharomyces and attempts to relate genetic, molecular biological, and biochemical studies of the processes of DNA repair and recombination.
Journal ArticleDOI

GINS maintains association of Cdc45 with MCM in replisome progression complexes at eukaryotic DNA replication forks

TL;DR: The GINS (go ichi ni san) complex allows the MCM (minichromosome maintenance) helicase to interact with key regulatory proteins in large replisome progression complexes (RPCs) that are assembled during initiation and disassembled at the end of S phase.
Journal ArticleDOI

DNA topoisomerase II and its growing repertoire of biological functions

TL;DR: Extensive biochemical and structural studies have provided detailed models of how TOP2 alters DNA structure, and recent molecular studies have greatly expanded knowledge of the biological contexts in which TOP2 functions, such as DNA replication, transcription and chromosome segregation.
Journal ArticleDOI

The yeast type I topoisomerase Top3 interacts with Sgs1, a DNA helicase homolog: a potential eukaryotic reverse gyrase.

TL;DR: In this paper, the SGS1 protein was identified as a slow growth suppressor that suppresses both the growth defect and the increased genomic instability of top-3 mutants in Saccharomyces cerevisiae, and it was shown that Sgs1 creates a deleterious topological substrate that Top3 preferentially resolves.
Journal ArticleDOI

Transcription generates positively and negatively supercoiled domains in the template.

TL;DR: The results indicate that the state of supercoiling of bacterial DNA is strongly modulated by transcription, and that DNA topoisomerases are normally involved in the elongation step of transcription.
References
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Journal ArticleDOI

A Film Detection Method for Tritium‐Labelled Proteins and Nucleic Acids in Polyacrylamide Gels

TL;DR: A simple method for detecting 3H in polyacrylamide gels by scintillation autography (fluorography) using X-ray film, which is ten times more sensitive than conventional autoradiography of isotopes with higher emission energies.
Journal ArticleDOI

DNA topoisomerase II is required at the time of mitosis in yeast

TL;DR: Five new temperature-sensitive DNA topoisomerase II mutations are constructed and their physiological consequences in yeast are analyzed, consistent with the hypothesis that the essential function of topoisomersase II is to allow the separation of intertwined chromosomal DNA molecules during mitosis.
Journal ArticleDOI

DNA topoisomerase II mutant of Saccharomyces cerevisiae: topoisomerase II is required for segregation of daughter molecules at the termination of DNA replication.

TL;DR: The results suggest that DNA topoisomerase II is necessary for the segregation of chromosomes at the termination of DNA replication.
Journal ArticleDOI

Isolation of type I and II DNA topoisomerase mutants from fission yeast: single and double mutants show different phenotypes in cell growth and chromatin organization.

TL;DR: The type II topoisomerase may have an essential role in the compaction and/or segregation of chromosomes during the nuclear division but also complement the defect of the type I enzyme whose major function is the maintenance of chromatin organization throughout the cell cycle.
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