The overall goal of the research proposed in this revised application is to study the cellular and viral processes involved in the initial stages of the infectious cycle of herpes simplex virus type 1 (HSV-1) and influenza A virus. As outlined by Dr. Helenius, the mission of a virus particle is to deliver the genome and accessory proteins into a host cell in a replication-competent form. The virus particle must enter the cytosol, and from the cytosol the genome and accessory proteins of HSV-1 and influenza viruses have to move into the nucleus for replication. The applicant proposes to particularly focus on the interaction of incoming HSV-1 capsids with elements of the cytoskeleton, and the mechanisms by which capsids are targeted to interact with nuclear pore complexes. With regard to influenza virus, the applicant proposes to study how the viral ribonucleoproteins (vRNPs) are transported into and out of the nucleus. Biological, biochemical and genetic approaches will be used to address the overall goals of the project. The specific goals of the proposed study are 1) to analyze the mechanisms by which incoming HSV-1 capsids interact with the peripheral cytoskeleton, associate with dynein, dynactin and microtubules, and how the capsids move to the nuclear pores; 2) to determine how HSV-1 capsids and influenza virus vRNPs interact with nuclear pores, and how the genome is transported through the pore; 3) to analyze the mechanism by which newly assembled influenza virus vRNPs are exported from the nucleus to the cytosol in infected cells. By addressing these largely neglected processes, the applicant hopes to unravel new principles and mechanisms relevant for the general understanding of virus-cell interactions. The results are also likely to provide new insight on the larger questions of cell tropism and pathogenesis. Moreover, the information may suggest novel antiviral strategies, and allow informed application of viruses as vehicles for gene transfer.

Agency
National Institute of Health (NIH)
Institute
National Institute of Allergy and Infectious Diseases (NIAID)
Type
Research Project (R01)
Project #
2R01AI018599-16A1
Application #
2402664
Study Section
Virology Study Section (VR)
Project Start
1982-01-01
Project End
1998-06-30
Budget Start
1997-07-01
Budget End
1998-06-30
Support Year
16
Fiscal Year
1997
Total Cost
Indirect Cost
Name
Yale University
Department
Anatomy/Cell Biology
Type
Schools of Medicine
DUNS #
082359691
City
New Haven
State
CT
Country
United States
Zip Code
06520
Ojala, P M; Sodeik, B; Ebersold, M W et al. (2000) Herpes simplex virus type 1 entry into host cells: reconstitution of capsid binding and uncoating at the nuclear pore complex in vitro. Mol Cell Biol 20:4922-31
Bui, M; Wills, E G; Helenius, A et al. (2000) Role of the influenza virus M1 protein in nuclear export of viral ribonucleoproteins. J Virol 74:1781-6
Alconada, A; Bauer, U; Sodeik, B et al. (1999) Intracellular traffic of herpes simplex virus glycoprotein gE: characterization of the sorting signals required for its trans-Golgi network localization. J Virol 73:377-87
Kann, M; Sodeik, B; Vlachou, A et al. (1999) Phosphorylation-dependent binding of hepatitis B virus core particles to the nuclear pore complex. J Cell Biol 145:45-55
Whittaker, G R; Helenius, A (1998) Nuclear import and export of viruses and virus genomes. Virology 246:1-23
Sodeik, B; Ebersold, M W; Helenius, A (1997) Microtubule-mediated transport of incoming herpes simplex virus 1 capsids to the nucleus. J Cell Biol 136:1007-21
Bui, M; Whittaker, G; Helenius, A (1996) Effect of M1 protein and low pH on nuclear transport of influenza virus ribonucleoproteins. J Virol 70:8391-401
Whittaker, G; Bui, M; Helenius, A (1996) Nuclear trafficking of influenza virus ribonuleoproteins in heterokaryons. J Virol 70:2743-56
Whittaker, G; Kemler, I; Helenius, A (1995) Hyperphosphorylation of mutant influenza virus matrix protein, M1, causes its retention in the nucleus. J Virol 69:439-45
Kemler, I; Whittaker, G; Helenius, A (1994) Nuclear import of microinjected influenza virus ribonucleoproteins. Virology 202:1028-33

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