Initiation of bacteriophage ø29 DNA packaging studied by optical tweezers manipulation of single DNA molecules

John Peter Rickgauer, Derek N. Fuller, Bo Hu, Shelley N Grimes, Paul J Jardine, Dwight L. Anderson, Douglas E. Smith

Research output: Chapter in Book/Report/Conference proceedingConference contribution

2 Scopus citations

Abstract

A key step in the life cycle of many viruses, including bacteriophages, adenoviruses, and herpesviruses, is the packaging of replicated viral genomes into pre-assembled proheads by the action of ATP-dependent portal motor complexes. Here we present a method that allows the initiation of packaging by single complexes to be studied using optical tweezers. A procedure is developed for assembling phage 029 prohead-motor complexes, which are demonstrated to bind and begin translocation of a target DNA molecule within only a few seconds. We show that the ø29 DNA terminal protein (gene product 3), which functions to prime DNA replication, also has a dramatic effect on packaging. The DNA tether length measured immediately after binding varied from ∼30-100% of the full length, yet shortened monotonically, indicating that packaging does not strictly begin at the terminal end of the DNA. Removal of the terminal protein eliminated this variability, causing packaging to initiate at or very near the end of the DNA. These findings, taken together with electron microscopy data, suggest that rather than simply threading into the portal, the motor captures and dynamically tensions a DNA loop, and that the function of the terminal protein is to load DNA segments on both sides of the loop junction onto separate DNA translocating units.

Original languageEnglish (US)
Title of host publicationOptical Trapping and Optical Micromanipulation III
DOIs
StatePublished - Dec 1 2006
EventOptical Trapping and Optical Micromanipulation III - San Diego, CA, United States
Duration: Aug 13 2006Aug 17 2006

Publication series

NameProceedings of SPIE - The International Society for Optical Engineering
Volume6326
ISSN (Print)0277-786X

Other

OtherOptical Trapping and Optical Micromanipulation III
CountryUnited States
CitySan Diego, CA
Period8/13/068/17/06

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Rickgauer, J. P., Fuller, D. N., Hu, B., Grimes, S. N., Jardine, P. J., Anderson, D. L., & Smith, D. E. (2006). Initiation of bacteriophage ø29 DNA packaging studied by optical tweezers manipulation of single DNA molecules. In Optical Trapping and Optical Micromanipulation III [632623] (Proceedings of SPIE - The International Society for Optical Engineering; Vol. 6326). https://doi.org/10.1117/12.682101