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

Abstract 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 in...

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Bibliographic Details
Published in:Nucleic acids research 2022-08, Vol.50 (15), p.8719-8732
Main Authors: 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
Format: Article
Language:English
Subjects:
Adenosine Triphosphatases - chemistry
Adenosine Triphosphatases - genetics
DNA Packaging
DNA, Viral - chemistry
DNA, Viral - genetics
Endodeoxyribonucleases - chemistry
Endodeoxyribonucleases - genetics
Molecular Biology
Viral Proteins - chemistry
Viral Proteins - genetics
Virus Assembly - genetics
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Summary:Abstract 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.
ISSN:0305-1048
1362-4962
DOI:10.1093/nar/gkac647