The assembly of influenza A virus involves the specific interaction of several viral components at a predetermined location within the virus-infected cell. The viral proteins involved in this process are key components in the generation of vaccines as well as in the adaption of virus to new hosts, but precise roles in the assembly process have yet to be ascribed or identified. The application of reverse genetics techniques and new technologies such as RNA interference allows for the precise dissection of the assembly process and the determination of precise functional domains important for virus assembly to take place. To fully understand the viral factors involved in influenza A virus assembly, a detailed molecular investigation of the domains of the M1 (matrix) and M2 proteins involved in virus assembly will be undertaken. The amino acid sequences in M1 and M2 that are important in targeting the proteins to sites of virus budding will be identified. The mechanism behind the anti-M2 antibody mediated inhibition of influenza A virus budding will be elucidated and specific interactions between the M1 protein and the M2 cytoplasmic tail will be characterized. Finally, the intracellular localization of M1 will be analyzed in cells infected with recombinant viruses bearing mutations in the HA and NA proteins that prevent their association with glycolipid rafts or the apical membrane. Recombinant viruses bearing specific mutations in one or several viral proteins will be generated and characterized for their sensitivity to anti-M2 antibodies, antiviral drug sensitivity and virulence. These studies should shed additional light on the mechanisms responsible for the generation of new, highly virulent influenza A viruses as well as further define the interactions that govern antibody and drug sensitivity.

Agency
National Institute of Health (NIH)
Institute
National Institute of Allergy and Infectious Diseases (NIAID)
Type
Research Project (R01)
Project #
5R01AI053629-04
Application #
6984804
Study Section
Experimental Virology Study Section (EVR)
Program Officer
Lacourciere, Karen A
Project Start
2003-06-15
Project End
2007-11-30
Budget Start
2005-12-01
Budget End
2006-11-30
Support Year
4
Fiscal Year
2006
Total Cost
$261,459
Indirect Cost
Name
Washington University
Department
Microbiology/Immun/Virology
Type
Schools of Medicine
DUNS #
068552207
City
Saint Louis
State
MO
Country
United States
Zip Code
63130
Stewart, Shaun M; Pekosz, Andrew (2012) The influenza C virus CM2 protein can alter intracellular pH, and its transmembrane domain can substitute for that of the influenza A virus M2 protein and support infectious virus production. J Virol 86:1277-81
Stewart, Shaun M; Pekosz, Andrew (2011) Mutations in the membrane-proximal region of the influenza A virus M2 protein cytoplasmic tail have modest effects on virus replication. J Virol 85:12179-87
Grantham, Michael L; Stewart, Shaun M; Lalime, Erin N et al. (2010) Tyrosines in the influenza A virus M2 protein cytoplasmic tail are critical for production of infectious virus particles. J Virol 84:8765-76
Stewart, Shaun M; Wu, Wai-Hong; Lalime, Erin N et al. (2010) The cholesterol recognition/interaction amino acid consensus motif of the influenza A virus M2 protein is not required for virus replication but contributes to virulence. Virology 405:530-8
Pekosz, Andrew; Newby, Celeste; Bose, Pulkit S et al. (2009) Sialic acid recognition is a key determinant of influenza A virus tropism in murine trachea epithelial cell cultures. Virology 386:61-7
Li, Yunsheng; Larrimer, Audrey; Curtiss, Teresa et al. (2009) Influenza virus assays based on virus-inducible reporter cell lines. Influenza Other Respir Viruses 3:241-51
Grantham, Michael L; Wu, Wai-Hong; Lalime, Erin N et al. (2009) Palmitoylation of the influenza A virus M2 protein is not required for virus replication in vitro but contributes to virus virulence. J Virol 83:8655-61
Pekosz, Andrew; Glass, Gregory E (2008) Emerging viral diseases. Md Med 9:11, 13-6
Wu, Wai-Hong; Pekosz, Andrew (2008) Extending the cytoplasmic tail of the influenza a virus M2 protein leads to reduced virus replication in vivo but not in vitro. J Virol 82:1059-63
Newby, Celeste M; Sabin, Leah; Pekosz, Andrew (2007) The RNA binding domain of influenza A virus NS1 protein affects secretion of tumor necrosis factor alpha, interleukin-6, and interferon in primary murine tracheal epithelial cells. J Virol 81:9469-80

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