The mechanism of DNA packaging for double-stranded DNA viruses will be studied in the Bacillus subtilis bacteriophage ?29, the most efficient in vitro viral packaging system known. Using an integrated genetic, biochemical and structural approach, we will characterize protein conformational change and movement in the transiently assembled packaging motor during DNA encapsidation. The mechanism of packaging in ?29 will serve as a model for animal virus packaging in the analogous herpesvirus and adenovirus systems, and aid in the search for new antiviral therapies. Due to similarities between the ?29 ATPase and other ring translocases, insights gained from the study of ?29 packaging will also provide insight into the basic principles of macromolecular motor function in higher organisms. To interrogate the mechanism of DNA packaging we will: complete and validate the atomic structure of the ?29 packaging motor using X-ray crystallography and cryoEM analysis (Aim 1); elucidate the mechanisms of force generation and intersubunit coordination during DNA translocation in a complex motor using integrated structural, genetic and biochemical approaches (Aim 2); and characterize the molecular events that culminate in termination of packaging using mutagenesis and biochemical analysis and cryoEM analysis (Aim 3).

Public Health Relevance

The assembly and movement of macromolecules in biological systems drive all aspects of health and disease at the cellular level. Study of the structure, assembly and function of the bacterial virus ?29 DNA packaging motor serves as a model for understanding these fundamental biological principles and can inform us about similar processes in higher organisms. The DNA packaging process of ?29 will also serve as a model for understanding similar events in the medically relevant herpesviruses and adenoviruses, thus providing targets for the development of new antiviral therapies.

Agency
National Institute of Health (NIH)
Institute
National Institute of General Medical Sciences (NIGMS)
Type
Research Project (R01)
Project #
2R01GM095516-05A1
Application #
9174559
Study Section
Prokaryotic Cell and Molecular Biology Study Section (PCMB)
Program Officer
Sakalian, Michael
Project Start
2011-09-30
Project End
2017-08-31
Budget Start
2016-09-30
Budget End
2017-08-31
Support Year
5
Fiscal Year
2016
Total Cost
$504,704
Indirect Cost
$95,599
Name
University of Minnesota Twin Cities
Department
Dentistry
Type
Schools of Dentistry
DUNS #
555917996
City
Minneapolis
State
MN
Country
United States
Zip Code
55455
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Mao, Huzhang; Saha, Mitul; Reyes-Aldrete, Emilio et al. (2016) Structural and Molecular Basis for Coordination in a Viral DNA Packaging Motor. Cell Rep 14:2017-2029
Zhao, Wei; Jardine, Paul J; Grimes, Shelley (2015) An RNA Domain Imparts Specificity and Selectivity to a Viral DNA Packaging Motor. J Virol 89:12457-66
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Zhao, Wei; Saha, Mitul; Ke, Ailong et al. (2012) A three-helix junction is the interface between two functional domains of prohead RNA in 29 DNA packaging. J Virol 86:11625-32
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