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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ftcdlib:oai:escholarship.org:ark:/13030/qt4w87691w 2023-09-05T13:15:22+02:00 Biochemical characterization of RecBCD enzyme from an Antarctic Pseudomonas species and identification of its cognate Chi (χ) sequence Pavankumar, Theetha L Sinha, Anurag K Ray, Malay K Spies, Maria e0197476 2018-01-01 application/pdf https://escholarship.org/uc/item/4w87691w unknown eScholarship, University of California qt4w87691w https://escholarship.org/uc/item/4w87691w public PLOS ONE, vol 13, iss 5 Genetics Underpinning research 1.1 Normal biological development and functioning Adenosine Triphosphatases Adenosine Triphosphate Antarctic Regions Base Sequence Cloning Molecular DNA Bacterial Exodeoxyribonuclease V Hydrolysis Magnesium Mutant Proteins Mutation Plasmids Pseudomonas Pseudomonas syringae Recombinant Fusion Proteins Substrate Specificity Temperature General Science & Technology article 2018 ftcdlib 2023-08-21T18:04:51Z 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. Article in Journal/Newspaper Antarc* Antarctic University of California: eScholarship Antarctic |
institution |
Open Polar |
collection |
University of California: eScholarship |
op_collection_id |
ftcdlib |
language |
unknown |
topic |
Genetics Underpinning research 1.1 Normal biological development and functioning Adenosine Triphosphatases Adenosine Triphosphate Antarctic Regions Base Sequence Cloning Molecular DNA Bacterial Exodeoxyribonuclease V Hydrolysis Magnesium Mutant Proteins Mutation Plasmids Pseudomonas Pseudomonas syringae Recombinant Fusion Proteins Substrate Specificity Temperature General Science & Technology |
spellingShingle |
Genetics Underpinning research 1.1 Normal biological development and functioning Adenosine Triphosphatases Adenosine Triphosphate Antarctic Regions Base Sequence Cloning Molecular DNA Bacterial Exodeoxyribonuclease V Hydrolysis Magnesium Mutant Proteins Mutation Plasmids Pseudomonas Pseudomonas syringae Recombinant Fusion Proteins Substrate Specificity Temperature General Science & Technology Pavankumar, Theetha L Sinha, Anurag K Ray, Malay K Biochemical characterization of RecBCD enzyme from an Antarctic Pseudomonas species and identification of its cognate Chi (χ) sequence |
topic_facet |
Genetics Underpinning research 1.1 Normal biological development and functioning Adenosine Triphosphatases Adenosine Triphosphate Antarctic Regions Base Sequence Cloning Molecular DNA Bacterial Exodeoxyribonuclease V Hydrolysis Magnesium Mutant Proteins Mutation Plasmids Pseudomonas Pseudomonas syringae Recombinant Fusion Proteins Substrate Specificity Temperature General Science & Technology |
description |
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. |
author2 |
Spies, Maria |
format |
Article in Journal/Newspaper |
author |
Pavankumar, Theetha L Sinha, Anurag K Ray, Malay K |
author_facet |
Pavankumar, Theetha L Sinha, Anurag K Ray, Malay K |
author_sort |
Pavankumar, Theetha L |
title |
Biochemical characterization of RecBCD enzyme from an Antarctic Pseudomonas species and identification of its cognate Chi (χ) sequence |
title_short |
Biochemical characterization of RecBCD enzyme from an Antarctic Pseudomonas species and identification of its cognate Chi (χ) sequence |
title_full |
Biochemical characterization of RecBCD enzyme from an Antarctic Pseudomonas species and identification of its cognate Chi (χ) sequence |
title_fullStr |
Biochemical characterization of RecBCD enzyme from an Antarctic Pseudomonas species and identification of its cognate Chi (χ) sequence |
title_full_unstemmed |
Biochemical characterization of RecBCD enzyme from an Antarctic Pseudomonas species and identification of its cognate Chi (χ) sequence |
title_sort |
biochemical characterization of recbcd enzyme from an antarctic pseudomonas species and identification of its cognate chi (χ) sequence |
publisher |
eScholarship, University of California |
publishDate |
2018 |
url |
https://escholarship.org/uc/item/4w87691w |
op_coverage |
e0197476 |
geographic |
Antarctic |
geographic_facet |
Antarctic |
genre |
Antarc* Antarctic |
genre_facet |
Antarc* Antarctic |
op_source |
PLOS ONE, vol 13, iss 5 |
op_relation |
qt4w87691w https://escholarship.org/uc/item/4w87691w |
op_rights |
public |
_version_ |
1776197163463737344 |