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Structural basis of DNA packaging by a ring-type ATPase from an archetypal viral system

Many essential cellular processes rely on substrate rotation or translocation by a multi-subunit, ring-type NTPase. A large number of double-stranded DNA viruses, including tailed bacteriophages and herpes viruses, use a homomeric ring ATPase to processively translocate viral genomic DNA into procap...

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Autores principales: Fung, Herman K H, Grimes, Shelley, Huet, Alexis, Duda, Robert L, Chechik, Maria, Gault, Joseph, Robinson, Carol V, Hendrix, Roger W, Jardine, Paul J, Conway, James F, Baumann, Christoph G, Antson, Alfred A
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Oxford University Press 2022
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC9410871/
https://www.ncbi.nlm.nih.gov/pubmed/35947691
http://dx.doi.org/10.1093/nar/gkac647
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author Fung, Herman K H
Grimes, Shelley
Huet, Alexis
Duda, Robert L
Chechik, Maria
Gault, Joseph
Robinson, Carol V
Hendrix, Roger W
Jardine, Paul J
Conway, James F
Baumann, Christoph G
Antson, Alfred A
author_facet Fung, Herman K H
Grimes, Shelley
Huet, Alexis
Duda, Robert L
Chechik, Maria
Gault, Joseph
Robinson, Carol V
Hendrix, Roger W
Jardine, Paul J
Conway, James F
Baumann, Christoph G
Antson, Alfred A
author_sort Fung, Herman K H
collection PubMed
description Many essential cellular processes rely on substrate rotation or translocation by a multi-subunit, ring-type NTPase. A large number of double-stranded DNA viruses, including tailed bacteriophages and herpes viruses, use a homomeric ring ATPase to processively translocate viral genomic DNA into procapsids during assembly. Our current understanding of viral DNA packaging comes from three archetypal bacteriophage systems: cos, pac and phi29. Detailed mechanistic understanding exists for pac and phi29, but not for cos. Here, we reconstituted in vitro a cos packaging system based on bacteriophage HK97 and provided a detailed biochemical and structural description. We used a photobleaching-based, single-molecule assay to determine the stoichiometry of the DNA-translocating ATPase large terminase. Crystal structures of the large terminase and DNA-recruiting small terminase, a first for a biochemically defined cos system, reveal mechanistic similarities between cos and pac systems. At the same time, mutational and biochemical analyses indicate a new regulatory mechanism for ATPase multimerization and coordination in the HK97 system. This work therefore establishes a framework for studying the evolutionary relationships between ATP-dependent DNA translocation machineries in double-stranded DNA viruses.
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spelling pubmed-94108712022-08-26 Structural basis of DNA packaging by a ring-type ATPase from an archetypal viral system Fung, Herman K H Grimes, Shelley Huet, Alexis Duda, Robert L Chechik, Maria Gault, Joseph Robinson, Carol V Hendrix, Roger W Jardine, Paul J Conway, James F Baumann, Christoph G Antson, Alfred A Nucleic Acids Res Molecular Biology Many essential cellular processes rely on substrate rotation or translocation by a multi-subunit, ring-type NTPase. A large number of double-stranded DNA viruses, including tailed bacteriophages and herpes viruses, use a homomeric ring ATPase to processively translocate viral genomic DNA into procapsids during assembly. Our current understanding of viral DNA packaging comes from three archetypal bacteriophage systems: cos, pac and phi29. Detailed mechanistic understanding exists for pac and phi29, but not for cos. Here, we reconstituted in vitro a cos packaging system based on bacteriophage HK97 and provided a detailed biochemical and structural description. We used a photobleaching-based, single-molecule assay to determine the stoichiometry of the DNA-translocating ATPase large terminase. Crystal structures of the large terminase and DNA-recruiting small terminase, a first for a biochemically defined cos system, reveal mechanistic similarities between cos and pac systems. At the same time, mutational and biochemical analyses indicate a new regulatory mechanism for ATPase multimerization and coordination in the HK97 system. This work therefore establishes a framework for studying the evolutionary relationships between ATP-dependent DNA translocation machineries in double-stranded DNA viruses. Oxford University Press 2022-08-10 /pmc/articles/PMC9410871/ /pubmed/35947691 http://dx.doi.org/10.1093/nar/gkac647 Text en © The Author(s) 2022. Published by Oxford University Press on behalf of Nucleic Acids Research. https://creativecommons.org/licenses/by/4.0/This is an Open Access article distributed under the terms of the Creative Commons Attribution License (https://creativecommons.org/licenses/by/4.0/), which permits unrestricted reuse, distribution, and reproduction in any medium, provided the original work is properly cited.
spellingShingle Molecular Biology
Fung, Herman K H
Grimes, Shelley
Huet, Alexis
Duda, Robert L
Chechik, Maria
Gault, Joseph
Robinson, Carol V
Hendrix, Roger W
Jardine, Paul J
Conway, James F
Baumann, Christoph G
Antson, Alfred A
Structural basis of DNA packaging by a ring-type ATPase from an archetypal viral system
title Structural basis of DNA packaging by a ring-type ATPase from an archetypal viral system
title_full Structural basis of DNA packaging by a ring-type ATPase from an archetypal viral system
title_fullStr Structural basis of DNA packaging by a ring-type ATPase from an archetypal viral system
title_full_unstemmed Structural basis of DNA packaging by a ring-type ATPase from an archetypal viral system
title_short Structural basis of DNA packaging by a ring-type ATPase from an archetypal viral system
title_sort structural basis of dna packaging by a ring-type atpase from an archetypal viral system
topic Molecular Biology
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC9410871/
https://www.ncbi.nlm.nih.gov/pubmed/35947691
http://dx.doi.org/10.1093/nar/gkac647
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