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

Herman K.H. Fung, Shelley N Grimes, Alexis Huet, Robert L. Duda, Maria Chechik, Joseph Gault, Carol V. Robinson, Roger W. Hendrix, Paul J. Jardine, James F. Conway, Christoph G. Baumann, Alfred A. Antson

Research output: Contribution to journalArticlepeer-review

3 Scopus citations

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.

Original languageEnglish (US)
Pages (from-to)8719-8732
Number of pages14
JournalNucleic acids research
Volume50
Issue number15
DOIs
StatePublished - Aug 26 2022

Bibliographical note

Publisher Copyright:
© The Author(s) 2022. Published by Oxford University Press on behalf of Nucleic Acids Research.

Fingerprint

Dive into the research topics of 'Structural basis of DNA packaging by a ring-type ATPase from an archetypal viral system'. Together they form a unique fingerprint.

Cite this