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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...
Autores principales: | , , , , , , , , , , , |
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Formato: | Online Artículo Texto |
Lenguaje: | English |
Publicado: |
Oxford University Press
2022
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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. |
format | Online Article Text |
id | pubmed-9410871 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2022 |
publisher | Oxford University Press |
record_format | MEDLINE/PubMed |
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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