The molecular mechanisms that are responsible for the expression and regulation of vaccinia virus (VV) late genes remain an enigma. This class of VV genes, which represents about half of the VV genetic potential, is expressed only after viral DNA synthesis has begun. VV late transcripts from any single locus are highly heterogeneous in size, apparently initiating from a number of distinct 5' sites and terminating randomly. The few VV late genes that have been mapped and sequenced to date, have revealed the conspicious absence of regulatory elements previously recognized as essential in other eukaryotic or prokaryotic systems, or even VV early genes. Thus, the identity of the VV late gene control regions and the viral proteins which presumably recognize them are unknown. Due to the complexity of the situation, one approach to unraveling these questions is to select a few VV late genes of interest, and to subject them to intense molecular biological scrutiny. Towards this end, a number of VV genes which participate in the late phase of VV replication [Alpha-amanitin resistance, ts17, and a tandem array of six non-coordinately expressed VV late genes] have recently been identified, mapped, and sequenced. The experiments seek to use the information obtained thus far as a basis for: 1) A comparative kinetic analysis of how and when these genes are expressed. 2) Preparation of immunological reagents sufficient to enable functional identification of the encoded gene products and how they participate in the viral life cycle. 3) Using directed-genetics, gene fusion, and marker rescue techniques to reveal the salient regulatory features of each gene. 4) Using footprinting and gel retardation methodologies to identify the viral and/or cellular proteins which interact with VV promoter and terminator regions. It is anticipated that such experiments will provide considerable insight into the mechanisms which VV employes to achieve the ordered expression of its complex developmental program within the infected cell. This information may facilitate the design and construction of future VV recombinant vaccine strains to be used for the prophylaxsis of infectious diseases.

Agency
National Institute of Health (NIH)
Institute
National Institute of Allergy and Infectious Diseases (NIAID)
Type
Research Project (R01)
Project #
5R01AI021335-06
Application #
3131339
Study Section
Experimental Virology Study Section (EVR)
Project Start
1983-12-01
Project End
1989-11-30
Budget Start
1988-12-01
Budget End
1989-11-30
Support Year
6
Fiscal Year
1989
Total Cost
Indirect Cost
Name
Oregon State University
Department
Type
Schools of Arts and Sciences
DUNS #
053599908
City
Corvallis
State
OR
Country
United States
Zip Code
97339
Alzhanova, Dina; Hruby, Dennis E (2007) A host cell membrane protein, golgin-97, is essential for poxvirus morphogenesis. Virology 362:421-7
Alzhanova, Dina; Hruby, Dennis E (2006) A trans-Golgi network resident protein, golgin-97, accumulates in viral factories and incorporates into virions during poxvirus infection. J Virol 80:11520-7
Blouch, Robert E; Byrd, Chelsea M; Hruby, Dennis E (2005) Importance of disulphide bonds for vaccinia virus L1R protein function. Virol J 2:91
Yoder, Jennifer D; Chen, Tsefang; Hruby, Dennis E (2004) Sequence-independent acylation of the vaccinia virus A-type inclusion protein. Biochemistry 43:8297-302
Chen, Tsefang F; Yoder, Jennifer D; Hruby, Dennis E (2004) Mass spectrometry analysis of synthetically myristoylated peptides. Eur J Mass Spectrom (Chichester, Eng) 10:501-8
Chen, Tsefang S; Yoder, Jennifer D; Hruby, Dennis E (2003) Preparation of a large hydrophobic protein for mass spectrometry analysis: vaccina virus ATI protein. Anal Biochem 315:277-80
Grosenbach, D W; Hansen, S G; Hruby, D E (2000) Identification and analysis of vaccinia virus palmitylproteins. Virology 275:193-206
Hansen, S G; Grosenbach, D W; Hruby, D E (1999) Analysis of the site occupancy constraints of primary amino acid sequences in the motif directing palmitylation of the vaccinia virus 37-kDa envelope protein. Virology 254:124-37
Martin, K H; Franke, C A; Hruby, D E (1999) Novel acylation of poxvirus A-type inclusion proteins. Virus Res 60:147-57
Grosenbach, D W; Hruby, D E (1998) Analysis of a vaccinia virus mutant expressing a nonpalmitylated form of p37, a mediator of virion envelopment. J Virol 72:5108-20

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