Abstract

The two cis-acting elements required for 3'-processing of pre-mRNA include the highly conserved AAUAAA hexamer and an amorphous U-or GU-rich downstream element. HIV-1 genome presents an interesting dilemma in that it selects 3'-poly (A) site while disregarding a canonical poly (A) site processing signal present at the 5'- end. An upstream element, some 76 nucleotides 3'- of AAUAAA hexamer, which is unique to the 3'poly (A) site, is required for efficient processing of the 3'- core poly (A). The upstream element serves to enhance 3'-processing by promoting stable binding of the processing factors, CPSF, (cleavage and polyadenylation specificity factor), at the core poly (A) site. The application aims to provide further insight into the sequence and structural elements of HIV-1 poly (A) site selection in order to design specific processing factor for the targeting of the 3'-processing machinery to the 5'- core poly (A) site. Such an accomplishment will block the synthesis of functional viral mRNAs. The PI proposes to develop strategies for targeting 3'-processing to multiple sites within the HIV-1 pre-mRNA, which will serve to restrict viral gene expression.

  Funding Agency

Agency
National Institute of Health (NIH)
Institute
National Institute of General Medical Sciences (NIGMS)
Type
Research Project (R01)
Project #
5R01GM046624-07
Application #
2684977
Study Section
AIDS and Related Research Study Section 3 (ARRC)
Project Start
1992-04-01
Project End
2000-03-31
Budget Start
1998-04-01
Budget End
1999-03-31
Support Year
7
Fiscal Year
1998
Total Cost
Indirect Cost

  Institution

Name
University of Vermont & St Agric College
Department
Microbiology/Immun/Virology
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