Biochemical characterization of RecBCD enzyme from an Antarctic Pseudomonas species and identification of its cognate Chi (χ) sequence
| Autor: | M. K. Ray, Anurag Kumar Sinha, Theetha L. Pavankumar |
|---|---|
| Přispěvatelé: | Spies, Maria |
| Rok vydání: | 2018 |
| Předmět: |
Adenosine Triphosphatase
0301 basic medicine Hydrolases Pseudomonas syringae lcsh:Medicine Pathology and Laboratory Medicine Biochemistry Substrate Specificity Endonuclease chemistry.chemical_compound Adenosine Triphosphate Medicine and Health Sciences Magnesium Cloning Molecular lcsh:Science Adenosine Triphosphatases Multidisciplinary biology Hydrolysis Bacterial Temperature Bacterial Pathogens Enzymes Nucleic acids Replication fork arrest Chemistry Medical Microbiology Physical Sciences Pathogens Research Article Chemical Elements Plasmids DNA Bacterial Exodeoxyribonuclease V Nucleases General Science & Technology DNA repair Recombinant Fusion Proteins 1.1 Normal biological development and functioning Protein subunit Nucleotide Sequencing Antarctic Regions DNA replication Research and Analysis Methods Microbiology 03 medical and health sciences Underpinning research Pseudomonas DNA-binding proteins Genetics Molecular Biology Techniques Sequencing Techniques Microbial Pathogens Molecular Biology RecBCD Nuclease Bacteria Base Sequence 030102 biochemistry & molecular biology lcsh:R Organisms Phosphatases Biology and Life Sciences Proteins Molecular DNA 030104 developmental biology chemistry Mutation Enzymology biology.protein bacteria Mutant Proteins lcsh:Q Cloning |
| Zdroj: | PLoS ONE, Vol 13, Iss 5, p e0197476 (2018) PloS one, vol 13, iss 5 PLoS ONE |
| ISSN: | 1932-6203 |
| DOI: | 10.1371/journal.pone.0197476 |
| Popis: | Pseudomonas syringaeLz4W RecBCD enzyme, RecBCDPs, is a trimeric protein complex comprised of RecC, RecB, and RecD subunits. RecBCD enzyme is essential forP. syringaegrowth at low temperature, and it protects cells from low temperature induced replication arrest. In this study, we show that the RecBCDPsenzyme displays distinct biochemical behaviors. UnlikeE. coliRecBCD enzyme, the RecD subunit is indispensable for RecBCDPsfunction. The RecD motor activity is essential for the Chi-like fragments production inP. syringae, highlighting a distinct role forP. syringaeRecD subunit in DNA repair and recombination process. Further, the ssDNA-dependent endonuclease activity is notably absent in RecBCDPsenzyme. Here, we demonstrate that the RecBCDPsenzyme 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 inP. syrinage; emphasizing the importance of DNA processing and recombination functions in rescuing low temperature induced replication fork arrest.Abbreviations:ATPAdenosine triphosphateDSBdouble-strand break‘ChiCrossover hotspot instigatorNi-NTANitrio tri-acetic acidTLCthin layer chromatographyMMCmitomycin CUV lightUltra violetABMAntarctic bacterial mediumLBLuria-Bertani medium |
| Databáze: | OpenAIRE |
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