All three subunits of RecBCD enzyme are essential for DNA repair and low-temperature growth in the Antarctic Pseudomonas syringae Lz4W.
BACKGROUND: The recD mutants of the Antarctic Pseudomonas syringae Lz4W are sensitive to DNA-damaging agents and fail to grow at 4 degrees C. Generally, RecD associates with two other proteins (RecB and RecC) to produce RecBCD enzyme, which is involved in homologous recombination and DNA repair in m...
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ftdoajarticles:oai:doaj.org/article:14bd6582d90a4b1986532d9e8832dc05 2023-05-15T13:47:39+02:00 All three subunits of RecBCD enzyme are essential for DNA repair and low-temperature growth in the Antarctic Pseudomonas syringae Lz4W. Theetha L Pavankumar Anurag K Sinha Malay K Ray 2010-01-01T00:00:00Z https://doi.org/10.1371/journal.pone.0009412 https://doaj.org/article/14bd6582d90a4b1986532d9e8832dc05 EN eng Public Library of Science (PLoS) http://europepmc.org/articles/PMC2828478?pdf=render https://doaj.org/toc/1932-6203 1932-6203 doi:10.1371/journal.pone.0009412 https://doaj.org/article/14bd6582d90a4b1986532d9e8832dc05 PLoS ONE, Vol 5, Iss 2, p e9412 (2010) Medicine R Science Q article 2010 ftdoajarticles https://doi.org/10.1371/journal.pone.0009412 2022-12-30T21:59:49Z BACKGROUND: The recD mutants of the Antarctic Pseudomonas syringae Lz4W are sensitive to DNA-damaging agents and fail to grow at 4 degrees C. Generally, RecD associates with two other proteins (RecB and RecC) to produce RecBCD enzyme, which is involved in homologous recombination and DNA repair in many bacteria, including Escherichia coli. However, RecD is not essential for DNA repair, nor does its deletion cause any growth defects in E. coli. Hence, the assessment of the P. syringae RecBCD pathway was imperative. METHODOLOGY/PRINCIPAL FINDINGS: Mutational analysis and genetic complementation studies were used to establish that the individual null-mutations of all three genes, recC, recB, and recD, or the deletion of whole recCBD operon of P. syringae, lead to growth inhibition at low temperature, and sensitivity to UV and mitomycin C. Viability of the mutant cells dropped drastically at 4 degrees C, and the mutants accumulated linear chromosomal DNA and shorter DNA fragments in higher amounts compared to 22 degrees C. Additional genetic data using the mutant RecBCD enzymes that were inactivated either in the ATPase active site of RecB (RecB(K29Q)) or RecD (RecD(K229Q)), or in the nuclease center of RecB (RecB(D1118A) and RecB(Delta nuc)) suggested that, while the nuclease activity of RecB is not so critical in vivo, the ATP-dependent functions of both RecB and RecD are essential. Surprisingly, E. coli recBCD or recBC alone on plasmid could complement the defects of the Delta recCBD strain of P. syringae. CONCLUSIONS/SIGNIFICANCE: All three subunits of the RecBCD(Ps) enzyme are essential for DNA repair and growth of P. syringae at low temperatures (4 degrees C). The RecD requirement is only a function of the RecBCD complex in the bacterium. The RecBCD pathway protects the Antarctic bacterium from cold-induced DNA damages, and is critically dependent on the helicase activities of both RecB and RecD subunits, but not on the nuclease of RecBCD(Ps) enzyme. Article in Journal/Newspaper Antarc* Antarctic Directory of Open Access Journals: DOAJ Articles Antarctic The Antarctic PLoS ONE 5 2 e9412 |
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Medicine R Science Q |
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Medicine R Science Q Theetha L Pavankumar Anurag K Sinha Malay K Ray All three subunits of RecBCD enzyme are essential for DNA repair and low-temperature growth in the Antarctic Pseudomonas syringae Lz4W. |
topic_facet |
Medicine R Science Q |
description |
BACKGROUND: The recD mutants of the Antarctic Pseudomonas syringae Lz4W are sensitive to DNA-damaging agents and fail to grow at 4 degrees C. Generally, RecD associates with two other proteins (RecB and RecC) to produce RecBCD enzyme, which is involved in homologous recombination and DNA repair in many bacteria, including Escherichia coli. However, RecD is not essential for DNA repair, nor does its deletion cause any growth defects in E. coli. Hence, the assessment of the P. syringae RecBCD pathway was imperative. METHODOLOGY/PRINCIPAL FINDINGS: Mutational analysis and genetic complementation studies were used to establish that the individual null-mutations of all three genes, recC, recB, and recD, or the deletion of whole recCBD operon of P. syringae, lead to growth inhibition at low temperature, and sensitivity to UV and mitomycin C. Viability of the mutant cells dropped drastically at 4 degrees C, and the mutants accumulated linear chromosomal DNA and shorter DNA fragments in higher amounts compared to 22 degrees C. Additional genetic data using the mutant RecBCD enzymes that were inactivated either in the ATPase active site of RecB (RecB(K29Q)) or RecD (RecD(K229Q)), or in the nuclease center of RecB (RecB(D1118A) and RecB(Delta nuc)) suggested that, while the nuclease activity of RecB is not so critical in vivo, the ATP-dependent functions of both RecB and RecD are essential. Surprisingly, E. coli recBCD or recBC alone on plasmid could complement the defects of the Delta recCBD strain of P. syringae. CONCLUSIONS/SIGNIFICANCE: All three subunits of the RecBCD(Ps) enzyme are essential for DNA repair and growth of P. syringae at low temperatures (4 degrees C). The RecD requirement is only a function of the RecBCD complex in the bacterium. The RecBCD pathway protects the Antarctic bacterium from cold-induced DNA damages, and is critically dependent on the helicase activities of both RecB and RecD subunits, but not on the nuclease of RecBCD(Ps) enzyme. |
format |
Article in Journal/Newspaper |
author |
Theetha L Pavankumar Anurag K Sinha Malay K Ray |
author_facet |
Theetha L Pavankumar Anurag K Sinha Malay K Ray |
author_sort |
Theetha L Pavankumar |
title |
All three subunits of RecBCD enzyme are essential for DNA repair and low-temperature growth in the Antarctic Pseudomonas syringae Lz4W. |
title_short |
All three subunits of RecBCD enzyme are essential for DNA repair and low-temperature growth in the Antarctic Pseudomonas syringae Lz4W. |
title_full |
All three subunits of RecBCD enzyme are essential for DNA repair and low-temperature growth in the Antarctic Pseudomonas syringae Lz4W. |
title_fullStr |
All three subunits of RecBCD enzyme are essential for DNA repair and low-temperature growth in the Antarctic Pseudomonas syringae Lz4W. |
title_full_unstemmed |
All three subunits of RecBCD enzyme are essential for DNA repair and low-temperature growth in the Antarctic Pseudomonas syringae Lz4W. |
title_sort |
all three subunits of recbcd enzyme are essential for dna repair and low-temperature growth in the antarctic pseudomonas syringae lz4w. |
publisher |
Public Library of Science (PLoS) |
publishDate |
2010 |
url |
https://doi.org/10.1371/journal.pone.0009412 https://doaj.org/article/14bd6582d90a4b1986532d9e8832dc05 |
geographic |
Antarctic The Antarctic |
geographic_facet |
Antarctic The Antarctic |
genre |
Antarc* Antarctic |
genre_facet |
Antarc* Antarctic |
op_source |
PLoS ONE, Vol 5, Iss 2, p e9412 (2010) |
op_relation |
http://europepmc.org/articles/PMC2828478?pdf=render https://doaj.org/toc/1932-6203 1932-6203 doi:10.1371/journal.pone.0009412 https://doaj.org/article/14bd6582d90a4b1986532d9e8832dc05 |
op_doi |
https://doi.org/10.1371/journal.pone.0009412 |
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PLoS ONE |
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5 |
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2 |
container_start_page |
e9412 |
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1766247587504979968 |