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On the viability of Escherichia coli cells lacking DNA topoisomerase I.

ABSTRACT: Manipulations of the DNA double helix during replication, transcription and other nucleic acid processing cause a change of DNA topology, which results in torsional stress. This stress is relaxed by DNA topoisomerases, a class of enzymes present in all domains of life. Negatively supercoiled DNA is relaxed by type IA topoisomerases that are widespread in bacteria, archaea and eukaryotes. In Escherichia coli there is conflicting data about viability of ?topA cells lacking topoisomerase I.In this study we sought to clarify whether E. coli cells lacking topoisomerase I are viable by using a plasmid-based lethality assay that allowed us to investigate the phenotype of ?topA cells without the presence of any compensatory mutations. Our results show that cells lacking topoisomerase I show an extreme growth defect and cannot be cultured without the accumulation of compensatory mutations. This growth defect can be partially suppressed by overexpression of topoisomerase III, the other type IA topoisomerase in E. coli, suggesting that the accumulation of torsional stress is, at least partially, responsible for the lethality of ?topA cells. The absence of RNase HI strongly exacerbates the phenotype of cells lacking topoisomerase I, which supports the idea that the processing of RNA:DNA hybrids is vitally important in ?topA cells. However, we did not observe suppression of the ?topA phenotype by increasing the level of R-loop processing enzymes, such as RNase HI or RecG.Our data show unambiguously that E. coli cells are not viable in the absence of DNA topoisomerase I without the presence of compensatory mutations. Furthermore, our data suggest that the accumulation of R-loops is not the primary reason for the severe growth defect of cells lacking topoisomerase I, which is in contrast to the current literature. Potential reasons for this discrepancy are discussed.

SUBMITTER: Stockum A

PROVIDER: S-EPMC3313902 | biostudies-other | 2012 Feb

REPOSITORIES: biostudies-other

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On the viability of Escherichia coli cells lacking DNA topoisomerase I.

Stockum Anna A Lloyd Robert G RG Rudolph Christian J CJ

BMC microbiology 20120228

<h4>Background</h4>Manipulations of the DNA double helix during replication, transcription and other nucleic acid processing cause a change of DNA topology, which results in torsional stress. This stress is relaxed by DNA topoisomerases, a class of enzymes present in all domains of life. Negatively supercoiled DNA is relaxed by type IA topoisomerases that are widespread in bacteria, archaea and eukaryotes. In Escherichia coli there is conflicting data about viability of ΔtopA cells lacking topoi ...[more]

PMID: 22373098

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Dataset Information

On the viability of Escherichia coli cells lacking DNA topoisomerase I.

Publications

On the viability of Escherichia coli cells lacking DNA topoisomerase I.

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