Abstract

The central goal of this project is to try to elucidate the biochemical basis of the regulation of HIV mRNA 3' processing. The long-term objective is to exploit the unique nature of the HIV poly(A) site processing signal to design an effective antiviral agent. The investigator proposes to identify the factor responsible for recognition of the HIV poly(A) site upstream element and the mechanism by which it enhances processing will be delineated.The subsequent cloning of this upstream factor and the characterization of its functional domains will permit the alteration of the RNA-binding specificity of the factor in order to facilitate recognition and subsequent processing of the HIV 5' core poly(A) site. A well-characterized in vitro processing system will be employed to probe the biochemical basis of the regulation of HIV mRNA 3' end formation. The investigator has five specific aims the first of which is to try to identify the processing factor that interacts with the HIV poly(A) site upstream element. Next, the author proposes to elucidate the mechanism by which the upstream factor functions to enhance processing at the HIV poly(A) site. Third, he will attempt to purify biochemically the HIV poly(A) site upstream factor and to isolate the corresponding cDNA clones. He also proposes to identify the domains of the upstream factor that mediate sequence recognition and function to enhance processing. Lastly, the author will try to develop a chimeric processing factor, and potential antiviral agent, to facilitate processing at the HIV 5' core poly(A) site.

  Funding Agency

Agency
National Institute of Health (NIH)
Institute
National Institute of General Medical Sciences (NIGMS)
Type
Research Project (R01)
Project #
5R01GM046624-02
Application #
3306056
Study Section
AIDS and Related Research Study Section 3 (ARRC)
Project Start
1992-04-01
Project End
1996-03-31
Budget Start
1993-04-01
Budget End
1994-03-31
Support Year
2
Fiscal Year
1993
Total Cost
Indirect Cost

  Institution

Name
University of Vermont & St Agric College
Department
Type
Schools of Medicine
DUNS #
066811191
City
Burlington
State
VT
Country
United States
Zip Code
05405

  Publications

Harakall, S A; Brandenburg, C A; Gilmartin, G A et al. (1998) Induction of multiple pituitary adenylate cyclase activating polypeptide (PACAP) transcripts through alternative cleavage and polyadenylation of proPACAP precursor mRNA. Ann N Y Acad Sci 865:367-74
Flaherty, S M; Fortes, P; Izaurralde, E et al. (1997) Participation of the nuclear cap binding complex in pre-mRNA 3' processing. Proc Natl Acad Sci U S A 94:11893-8
Gilmartin, G M; Hung, S L; DeZazzo, J D et al. (1996) Sequences regulating poly(A) site selection within the adenovirus major late transcription unit influence the interaction of constitutive processing factors with the pre-mRNA. J Virol 70:1775-83
Graveley, B R; Gilmartin, G M (1996) A common mechanism for the enhancement of mRNA 3' processing by U3 sequences in two distantly related lentiviruses. J Virol 70:1612-7
Graveley, B R; Fleming, E S; Gilmartin, G M (1996) Restoration of both structure and function to a defective poly(A) site by in vitro selection. J Biol Chem 271:33654-63
Graveley, B R; Fleming, E S; Gilmartin, G M (1996) RNA structure is a critical determinant of poly(A) site recognition by cleavage and polyadenylation specificity factor. Mol Cell Biol 16:4942-51
Gilmartin, G M; Fleming, E S; Oetjen, J et al. (1995) CPSF recognition of an HIV-1 mRNA 3'-processing enhancer: multiple sequence contacts involved in poly(A) site definition. Genes Dev 9:72-83
Gilmartin, G M; Fleming, E S; Oetjen, J (1992) Activation of HIV-1 pre-mRNA 3' processing in vitro requires both an upstream element and TAR. EMBO J 11:4419-28