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Structural studies reveal a ring-shaped architecture of deep-sea vent phage NrS-1 polymerase

NrS-1 is the first known phage that can infect Epsilonproteobacteria, one of the predominant primary producers in the deep-sea hydrothermal vent ecosystems. NrS-1 polymerase is a multidomain enzyme and is one key component of the phage replisome. The N-terminal Prim/Pol and HBD domains are responsib...

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Autores principales: Chen, Xi, Su, Shichen, Chen, Yiqing, Gao, Yanqing, Li, Yangyang, Shao, Zhiwei, Zhang, Yixi, Shao, Qiyuan, Liu, Hehua, Li, Jixi, Ma, Jinbiao, Gan, Jianhua
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Oxford University Press 2020
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC7102993/
https://www.ncbi.nlm.nih.gov/pubmed/32016421
http://dx.doi.org/10.1093/nar/gkaa071
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author Chen, Xi
Su, Shichen
Chen, Yiqing
Gao, Yanqing
Li, Yangyang
Shao, Zhiwei
Zhang, Yixi
Shao, Qiyuan
Liu, Hehua
Li, Jixi
Ma, Jinbiao
Gan, Jianhua
author_facet Chen, Xi
Su, Shichen
Chen, Yiqing
Gao, Yanqing
Li, Yangyang
Shao, Zhiwei
Zhang, Yixi
Shao, Qiyuan
Liu, Hehua
Li, Jixi
Ma, Jinbiao
Gan, Jianhua
author_sort Chen, Xi
collection PubMed
description NrS-1 is the first known phage that can infect Epsilonproteobacteria, one of the predominant primary producers in the deep-sea hydrothermal vent ecosystems. NrS-1 polymerase is a multidomain enzyme and is one key component of the phage replisome. The N-terminal Prim/Pol and HBD domains are responsible for DNA polymerization and de novo primer synthesis activities of NrS-1 polymerase. However, the structure and function of the C-terminus (CTR) of NrS-1 polymerase are poorly understood. Here, we report two crystal structures, showing that NrS-1 CTR adopts one unique hexameric ring-shaped conformation. Although the central helicase domain of NrS-1 CTR shares structural similarity with the superfamily III helicases, the folds of the Head and Tail domains are completely novel. Via mutagenesis and in vitro biochemical analysis, we identified many residues important for the helicase and polymerization activities of NrS-1 polymerase. In addition to NrS-1 polymerase, our study may also help us identify and understand the functions of multidomain polymerases expressed by many NrS-1 related phages.
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spelling pubmed-71029932020-04-02 Structural studies reveal a ring-shaped architecture of deep-sea vent phage NrS-1 polymerase Chen, Xi Su, Shichen Chen, Yiqing Gao, Yanqing Li, Yangyang Shao, Zhiwei Zhang, Yixi Shao, Qiyuan Liu, Hehua Li, Jixi Ma, Jinbiao Gan, Jianhua Nucleic Acids Res Structural Biology NrS-1 is the first known phage that can infect Epsilonproteobacteria, one of the predominant primary producers in the deep-sea hydrothermal vent ecosystems. NrS-1 polymerase is a multidomain enzyme and is one key component of the phage replisome. The N-terminal Prim/Pol and HBD domains are responsible for DNA polymerization and de novo primer synthesis activities of NrS-1 polymerase. However, the structure and function of the C-terminus (CTR) of NrS-1 polymerase are poorly understood. Here, we report two crystal structures, showing that NrS-1 CTR adopts one unique hexameric ring-shaped conformation. Although the central helicase domain of NrS-1 CTR shares structural similarity with the superfamily III helicases, the folds of the Head and Tail domains are completely novel. Via mutagenesis and in vitro biochemical analysis, we identified many residues important for the helicase and polymerization activities of NrS-1 polymerase. In addition to NrS-1 polymerase, our study may also help us identify and understand the functions of multidomain polymerases expressed by many NrS-1 related phages. Oxford University Press 2020-04-06 2020-02-04 /pmc/articles/PMC7102993/ /pubmed/32016421 http://dx.doi.org/10.1093/nar/gkaa071 Text en © The Author(s) 2020. Published by Oxford University Press on behalf of Nucleic Acids Research. http://creativecommons.org/licenses/by/4.0/ This is an Open Access article distributed under the terms of the Creative Commons Attribution License (http://creativecommons.org/licenses/by/4.0/), which permits unrestricted reuse, distribution, and reproduction in any medium, provided the original work is properly cited.
spellingShingle Structural Biology
Chen, Xi
Su, Shichen
Chen, Yiqing
Gao, Yanqing
Li, Yangyang
Shao, Zhiwei
Zhang, Yixi
Shao, Qiyuan
Liu, Hehua
Li, Jixi
Ma, Jinbiao
Gan, Jianhua
Structural studies reveal a ring-shaped architecture of deep-sea vent phage NrS-1 polymerase
title Structural studies reveal a ring-shaped architecture of deep-sea vent phage NrS-1 polymerase
title_full Structural studies reveal a ring-shaped architecture of deep-sea vent phage NrS-1 polymerase
title_fullStr Structural studies reveal a ring-shaped architecture of deep-sea vent phage NrS-1 polymerase
title_full_unstemmed Structural studies reveal a ring-shaped architecture of deep-sea vent phage NrS-1 polymerase
title_short Structural studies reveal a ring-shaped architecture of deep-sea vent phage NrS-1 polymerase
title_sort structural studies reveal a ring-shaped architecture of deep-sea vent phage nrs-1 polymerase
topic Structural Biology
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC7102993/
https://www.ncbi.nlm.nih.gov/pubmed/32016421
http://dx.doi.org/10.1093/nar/gkaa071
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