Poliovirus provides an extraordinary opportunity to understand a complex biological system at the molecular level. The results of over forty years of characterization have established poliovirus as paradigm for understanding the molecular biology of simple viruses and the nature of their interactions with their hosts. In the past ten years the introduction of genome sequencing methods and the availability of infectious clones of the virus have provided the ability to map the determinants of biological properties of the virus to specific locations in the viral genome. The availability of high resolution structures of poliovirus (which have been solved under the auspices of this grant) has permitted a number these properties to be interpreted in the context of the three-dimensional structure of the virion, and has provided significant insights into the role of conformational transitions in the life cycle of the virus. In this application we propose to continue our structural studies of poliovirus in order to further probe the relationship between its structure and biological functions. Crystallographic studies will be extended to include several intermediates in the assembly process (protomers, pentamers empty capsids, and provirions), a cell entry intermediate, chimeric viruses expressing antigenic determinants of heterologous pathogens (including HIV), and virus strains and variants with specific alterations in assembly, host-range, neurovirulence, and receptor interactions. Crystallographic studies will also be extended to include Fab fragments of a monoclonal antibody, C3 (which is primarily directed against a single loop in VP1 and binds peptides from this loop) and soluble fragments of the poliovirus receptor. The targets have been chosen to answer specific biological questions, including the role of conformational changes in the assembly and cell entry of the virus, the nature of the interactions of the virus with its receptor and with the host's immune system, and the determinants of host-range and pathogenesis. The crystallographic analyses will be carried out in collaboration with a number other research groups in order to provide a direct interplay between structural molecular genetic studies.

Agency
National Institute of Health (NIH)
Institute
National Institute of Allergy and Infectious Diseases (NIAID)
Type
Method to Extend Research in Time (MERIT) Award (R37)
Project #
5R37AI020566-17
Application #
2855931
Study Section
Special Emphasis Panel (NSS)
Program Officer
Meegan, James M
Project Start
1983-12-01
Project End
2001-12-31
Budget Start
1999-01-01
Budget End
1999-12-31
Support Year
17
Fiscal Year
1999
Total Cost
Indirect Cost
Name
Harvard University
Department
Biochemistry
Type
Schools of Medicine
DUNS #
082359691
City
Boston
State
MA
Country
United States
Zip Code
02115
Zhao, Zhao; Zhang, Meng; Hogle, James M et al. (2018) DNA-Corralled Nanodiscs for the Structural and Functional Characterization of Membrane Proteins and Viral Entry. J Am Chem Soc 140:10639-10643
Nasr, Mahmoud L; Baptista, Diego; Strauss, Mike et al. (2017) Covalently circularized nanodiscs for studying membrane proteins and viral entry. Nat Methods 14:49-52
Strauss, Mike; Schotte, Lise; Karunatilaka, Krishanthi S et al. (2017) Cryo-electron Microscopy Structures of Expanded Poliovirus with VHHs Sample the Conformational Repertoire of the Expanded State. J Virol 91:
Strauss, Mike; Schotte, Lise; Thys, Bert et al. (2016) Five of Five VHHs Neutralizing Poliovirus Bind the Receptor-Binding Site. J Virol 90:3496-505
Schotte, Lise; Thys, Bert; Strauss, Mike et al. (2015) Characterization of Poliovirus Neutralization Escape Mutants of Single-Domain Antibody Fragments (VHHs). Antimicrob Agents Chemother 59:4695-706
Strauss, Mike; Filman, David J; Belnap, David M et al. (2015) Nectin-like interactions between poliovirus and its receptor trigger conformational changes associated with cell entry. J Virol 89:4143-57
Schotte, Lise; Strauss, Mike; Thys, Bert et al. (2014) Mechanism of action and capsid-stabilizing properties of VHHs with an in vitro antipolioviral activity. J Virol 88:4403-13
Butan, Carmen; Filman, David J; Hogle, James M (2014) Cryo-electron microscopy reconstruction shows poliovirus 135S particles poised for membrane interaction and RNA release. J Virol 88:1758-70
Strauss, Mike; Levy, Hazel C; Bostina, Mihnea et al. (2013) RNA transfer from poliovirus 135S particles across membranes is mediated by long umbilical connectors. J Virol 87:3903-14
Levy, Hazel C; Bostina, Mihnea; Filman, David J et al. (2010) Catching a virus in the act of RNA release: a novel poliovirus uncoating intermediate characterized by cryo-electron microscopy. J Virol 84:4426-41

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