The primary goal is to determine the mechanism by which lipid containing viruses assemble and bud from the cell membrane. This project deals with the main structural component of influenza virus, the Ml protein and has two purposes. First, the exact association of Ml with the cell lipid and the interactions of Ml with the other viral proteins that leads to virus formation will be determined. Second, the importance of phosphorylation of Ml in virus assembly will be investigated by establishing the quantify of phosphate, the site of phosphorylation and determining if there is conservation of phosphorylation of the Ml in groups B and C influenza viruses. These studies will involve the use of chemical cross-linking reagents of varying lengths and varying types, isolation of components by immunoprecipitation, isolation of subcellular fractions on gradients and immunofluorescent techniques on whole cells. Assays will be used for quantitating phosphate. Proteolytic enzymes and chemical agents will be used to cleave Ml. Chromatographic procedures will be used to separate peptides which will be analyzed for amino acid content and sequence. A recombinant virus containing a clone of Ml will be used to study the pathway and association of Ml in the cell and its phosphorylation in the absence of other proteins.

Agency
National Institute of Health (NIH)
Institute
National Institute of Allergy and Infectious Diseases (NIAID)
Type
Research Project (R01)
Project #
5R01AI023446-02
Application #
3135519
Study Section
Experimental Virology Study Section (EVR)
Project Start
1986-09-01
Project End
1988-11-30
Budget Start
1987-09-01
Budget End
1988-11-30
Support Year
2
Fiscal Year
1987
Total Cost
Indirect Cost
Name
New York College of Podiatric Medicine
Department
Type
Colleges of Podiatric Medicine
DUNS #
City
New York
State
NY
Country
United States
Zip Code
10035
Morrison, D C; Lei, M G; Chen, T Y et al. (1992) Identification and characterization of mammalian cell membrane receptors for LPS-endotoxin. Adv Exp Med Biol 319:23-9
Perera, P Y; Chan, T Y; Morrison, D C et al. (1992) Detection and analysis of the 80-kd lipopolysaccharide receptor in macrophages derived from Lpsn and Lpsd mice. J Leukoc Biol 51:501-6