Influenza viruses orthomyxoviruses, a major group of human and animal pathogens, are a group of segmented enveloped negative strand RNA viruses. These viruses assemble and bud at the level of plasma membrane, specifically on the apical side of the polarized epithelial cells. The long term goal of this project define the processes of viral assembly and budding. The specific objectives in the proposed project are to examine the functions of M1 and NA in the assembly and budding processes. Since influenza virus replication occurs in the cell's nucleus and budding takes place at the plasma membrane, how the viral RNP gets in and out of the nucleus and how dissociation and association of M1 facilitates the nuclear entry and exit of vRNP will be examined. Proposed experiments will determine if M1 becomes dissociated from the viral RNP during uncoating and if the preexisting M1 will interfere with the nuclear transport of the incoming vRNP. In addition, these experiments will determine if vRNP made in the absence of M1 is exported out of the nucleus into the cytoplasm or if M1 is needed for vRNP's exit from the nucleus. Furthermore the domains of M1 involved in interaction with vRNP will be delineated. Since M1 is juxtaposed between the viral envelope and RNP, the protein-protein interactions between M1 and the transmembrane viral proteins like HA, NA or M2 and the domains of each of these proteins involved in the interactions will be defined. Since viral particles bud from the apical domain of plasma membrane in polarized cells, structural features present in these transmembrane proteins, particularly in NA, responsible for targeting the protein to the apical plasma membrane, will be determined. NA is a type II membrane protein. Little is known about the sorting signals for polarized transport of type II protein in general and NA in particular. Chimeric constructions and deletions and mutations will be used to define sorting signals of NA. Experiments proposed here will elucidate the steps involved in uncoating as well as assembly and budding and will be useful in designing the antiviral agents to block these steps in viral replication.

Agency
National Institute of Health (NIH)
Institute
National Institute of Allergy and Infectious Diseases (NIAID)
Type
Research Project (R01)
Project #
5R01AI016348-15
Application #
2060356
Study Section
Virology Study Section (VR)
Project Start
1980-05-01
Project End
1999-03-31
Budget Start
1995-04-01
Budget End
1996-03-31
Support Year
15
Fiscal Year
1995
Total Cost
Indirect Cost
Name
University of California Los Angeles
Department
Microbiology/Immun/Virology
Type
Schools of Medicine
DUNS #
119132785
City
Los Angeles
State
CA
Country
United States
Zip Code
90095
Nayak, Debi P; Balogun, Rilwan A; Yamada, Hiroshi et al. (2009) Influenza virus morphogenesis and budding. Virus Res 143:147-61
Barman, Subrata; Nayak, Debi P (2007) Lipid raft disruption by cholesterol depletion enhances influenza A virus budding from MDCK cells. J Virol 81:12169-78
Hui, Eric Ka-Wai; Smee, Donald F; Wong, Min-Hui et al. (2006) Mutations in influenza virus M1 CCHH, the putative zinc finger motif, cause attenuation in mice and protect mice against lethal influenza virus infection. J Virol 80:5697-707
Hui, Eric Ka-Wai; Barman, Subrata; Tang, Dominic Ho-Ping et al. (2006) YRKL sequence of influenza virus M1 functions as the L domain motif and interacts with VPS28 and Cdc42. J Virol 80:2291-308
Hui, Eric Ka-Wai; Yap, Ee Ming; An, Dong Sung et al. (2004) Inhibition of influenza virus matrix (M1) protein expression and virus replication by U6 promoter-driven and lentivirus-mediated delivery of siRNA. J Gen Virol 85:1877-84
Nayak, Debi P; Hui, Eric Ka-Wai; Barman, Subrata (2004) Assembly and budding of influenza virus. Virus Res 106:147-65
Nayak, Debi P; Hui, Eric K W (2004) The role of lipid microdomains in virus biology. Subcell Biochem 37:443-91
Barman, Subrata; Adhikary, Lopa; Chakrabarti, Alok K et al. (2004) Role of transmembrane domain and cytoplasmic tail amino acid sequences of influenza a virus neuraminidase in raft association and virus budding. J Virol 78:5258-69
Hui, Eric Ka-Wai; Ralston, Katherine; Judd, Amrit K et al. (2003) Conserved cysteine and histidine residues in the putative zinc finger motif of the influenza A virus M1 protein are not critical for influenza virus replication. J Gen Virol 84:3105-13
Barman, Subrata; Adhikary, Lopa; Kawaoka, Yoshihiro et al. (2003) Influenza A virus hemagglutinin containing basolateral localization signal does not alter the apical budding of a recombinant influenza A virus in polarized MDCK cells. Virology 305:138-52

Showing the most recent 10 out of 43 publications