Human (HIV) and bovine (BIV) immunodeficiency viruses exploit host cyclin T1 proteins to enhance viral transcription using interactions between cyclin T1, viral Tat proteins, and the TAR RNA hairpins located on viral transcripts. The two viruses have evolved distinct mechanisms for cyclin T1 recognition; however, these differences are not well understood in molecular detail. Biochemical and biophysical experiments will be used to characterize the domains of cyclin T1 that contain both the canonical cyclin fold and the binding region for the Tat:TAR complex. This includes purification of the human and bovine cyclin domains and assessment of protein folding by both spectroscopic assays, and proteolytic mapping. Additionally, individual contributions of cyclin, Tat, and TAR molecules to the formation of the protein-RNA complexes will be assayed through a series of titration experiments and proteolytic mapping. Finally, optimizing the conditions to generate correctly folded complexes will facilitate attempts at growing crystals of the complexes. The molecular details uncovered by these studies are expected to provide new information about transcription control mechanisms and may aid in the search for new therapies to treat viral disease and cyclin related cancers.

Agency
National Institute of Health (NIH)
Institute
National Institute of General Medical Sciences (NIGMS)
Type
Postdoctoral Individual National Research Service Award (F32)
Project #
5F32GM066542-03
Application #
6844709
Study Section
Special Emphasis Panel (ZRG1-F09 (20))
Program Officer
Portnoy, Matthew
Project Start
2003-02-01
Project End
2005-03-04
Budget Start
2005-02-01
Budget End
2005-03-04
Support Year
3
Fiscal Year
2005
Total Cost
$7,352
Indirect Cost
Name
Scripps Research Institute
Department
Type
DUNS #
781613492
City
La Jolla
State
CA
Country
United States
Zip Code
92037
Edgcomb, Stephen P; Aschrafi, Angelique; Kompfner, Elizabeth et al. (2008) Protein structure and oligomerization are important for the formation of export-competent HIV-1 Rev-RRE complexes. Protein Sci 17:420-30