Biochemical characterization of RecBCD enzyme from an Antarctic Pseudomonas species and identification of its cognate Chi (χ) sequence

Pseudomonas syringae Lz4W RecBCD enzyme, RecBCDPs, is a trimeric protein complex comprised of RecC, RecB, and RecD subunits. RecBCD enzyme is essential for P. syringae growth at low temperature, and it protects cells from low temperature induced replication arrest. In this study, we show that the Re...

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Bibliographic Details
Published in:PloS one 2018-05, Vol.13 (5), p.e0197476-e0197476
Main Authors: Pavankumar, Theetha L, Sinha, Anurag K, Ray, Malay K
Format: Article
Language:English
Subjects:
Analysis
Base sequence
Biology and Life Sciences
Chromatography
Deoxyribonucleic acid
DNA
DNA repair
E coli
Enzymes
Escherichia coli
Genetic aspects
Genomes
Low temperature
Medicine and Health Sciences
Molecular biology
Motor activity
Mutants
Nitrates
Nuclease
Nucleotide sequence
Physical Sciences
Proteins
Pseudomonas
Recombination
Replication
Research and Analysis Methods
Temperature effects
Translocation
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Summary:Pseudomonas syringae Lz4W RecBCD enzyme, RecBCDPs, is a trimeric protein complex comprised of RecC, RecB, and RecD subunits. RecBCD enzyme is essential for P. syringae growth at low temperature, and it protects cells from low temperature induced replication arrest. In this study, we show that the RecBCDPs enzyme displays distinct biochemical behaviors. Unlike E. coli RecBCD enzyme, the RecD subunit is indispensable for RecBCDPs function. The RecD motor activity is essential for the Chi-like fragments production in P. syringae, highlighting a distinct role for P. syringae RecD subunit in DNA repair and recombination process. Here, we demonstrate that the RecBCDPs enzyme recognizes a unique octameric DNA sequence, 5'-GCTGGCGC-3' (ChiPs) that attenuates nuclease activity of the enzyme when it enters dsDNA from the 3'-end. We propose that the reduced translocation activities manifested by motor-defective mutants cause cold sensitivity in P. syrinage; emphasizing the importance of DNA processing and recombination functions in rescuing low temperature induced replication fork arrest.
ISSN:1932-6203
1932-6203
DOI:10.1371/journal.pone.0197476