Analysis of topoisomerase function in bacterial replication fork movement: Use of DNA microarrays

Arkady B. Khodursky; Brian J. Peter; Molly B. Schmid; Joseph DeRisi; David Botstein; Patrick O. Brown; Nicholas R. Cozzarelli

doi:10.1073/pnas.97.17.9419

Analysis of topoisomerase function in bacterial replication fork movement: Use of DNA microarrays

Arkady B. Khodursky, Brian J. Peter, Molly B. Schmid, Joseph DeRisi, David Botstein, Patrick O. Brown, Nicholas R. Cozzarelli

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164 Scopus citations

Abstract

We used DNA microarrays of the Escherichia coli genome to trace the progression of chromosomal replication forks in synchronized cells. We found that both DNA gyrase and topoisomerase IV (topo IV) promote replication fork progression. When both enzymes were inhibited, the replication fork stopped rapidly. The elongation rate with topo IV alone was 1/3 of normal. Genetic data confirmed and extended these results. Inactivation of gyrase alone caused a slow stop of replication. Topo IV activity was sufficient to prevent accumulation of (+) supercoils in plasmid DNA in vivo, suggesting that topo IV can promote replication by removing (+) supercoils in front of the chromosomal fork.

Original language	English (US)
Pages (from-to)	9419-9424
Number of pages	6
Journal	Proceedings of the National Academy of Sciences of the United States of America
Volume	97
Issue number	17
DOIs	https://doi.org/10.1073/pnas.97.17.9419
State	Published - Aug 15 2000
Externally published	Yes

Keywords

Coumarins
Positive supercoiling
Topo IV

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10.1073/pnas.97.17.9419

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abstract = "We used DNA microarrays of the Escherichia coli genome to trace the progression of chromosomal replication forks in synchronized cells. We found that both DNA gyrase and topoisomerase IV (topo IV) promote replication fork progression. When both enzymes were inhibited, the replication fork stopped rapidly. The elongation rate with topo IV alone was 1/3 of normal. Genetic data confirmed and extended these results. Inactivation of gyrase alone caused a slow stop of replication. Topo IV activity was sufficient to prevent accumulation of (+) supercoils in plasmid DNA in vivo, suggesting that topo IV can promote replication by removing (+) supercoils in front of the chromosomal fork.",

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T1 - Analysis of topoisomerase function in bacterial replication fork movement

T2 - Use of DNA microarrays

AU - Khodursky, Arkady B.

AU - Peter, Brian J.

AU - Schmid, Molly B.

AU - DeRisi, Joseph

AU - Botstein, David

AU - Brown, Patrick O.

AU - Cozzarelli, Nicholas R.

PY - 2000/8/15

Y1 - 2000/8/15

N2 - We used DNA microarrays of the Escherichia coli genome to trace the progression of chromosomal replication forks in synchronized cells. We found that both DNA gyrase and topoisomerase IV (topo IV) promote replication fork progression. When both enzymes were inhibited, the replication fork stopped rapidly. The elongation rate with topo IV alone was 1/3 of normal. Genetic data confirmed and extended these results. Inactivation of gyrase alone caused a slow stop of replication. Topo IV activity was sufficient to prevent accumulation of (+) supercoils in plasmid DNA in vivo, suggesting that topo IV can promote replication by removing (+) supercoils in front of the chromosomal fork.

AB - We used DNA microarrays of the Escherichia coli genome to trace the progression of chromosomal replication forks in synchronized cells. We found that both DNA gyrase and topoisomerase IV (topo IV) promote replication fork progression. When both enzymes were inhibited, the replication fork stopped rapidly. The elongation rate with topo IV alone was 1/3 of normal. Genetic data confirmed and extended these results. Inactivation of gyrase alone caused a slow stop of replication. Topo IV activity was sufficient to prevent accumulation of (+) supercoils in plasmid DNA in vivo, suggesting that topo IV can promote replication by removing (+) supercoils in front of the chromosomal fork.

KW - Coumarins

KW - Positive supercoiling

KW - Topo IV

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Analysis of topoisomerase function in bacterial replication fork movement: Use of DNA microarrays

Abstract

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