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

Herman K H Fung; Shelley 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

doi:10.1093/nar/gkac647

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

Herman K H Fung, Shelley 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 journal › Article › peer-review

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 language	English
Article number	gkac647
Number of pages	14
Journal	Nucleic Acids Research
Early online date	10 Aug 2022
DOIs	https://doi.org/10.1093/nar/gkac647
Publication status	E-pub ahead of print - 10 Aug 2022

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10.1093/nar/gkac647Licence: CC BY

Structural basis of DNA packaging by a ring-type ATPase from an archetypal viral system
© The Author(s) 2022.
Final published version, 3.13 MBLicence: CC BY

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Fung, H. K. H., Grimes, S., Huet, A., Duda, R. L., Chechik, M., Gault, J., Robinson, C. V., Hendrix, R. W., Jardine, P. J., Conway, J. F., Baumann, C. G., & Antson, A. A. (2022). Structural basis of DNA packaging by a ring-type ATPase from an archetypal viral system. Nucleic Acids Research, Article gkac647. Advance online publication. https://doi.org/10.1093/nar/gkac647

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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.",

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AU - Gault, Joseph

AU - Robinson, Carol V

AU - Hendrix, Roger W

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AU - Conway, James F

AU - Baumann, Christoph G

AU - Antson, Alfred A

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