This is a proposal to create new aptamers to reverse transcriptase via SELEX and to characterize the interaction of these aptamers in detail. Novel approaches to generate chimeric aptamers will be used in the proposed studies and the interaction of these aptamers with RT will be evaluated through the use of several complementary biochemical and physical assays. In addition, experiments will be carried out to see what kinds of RT resistant mutants arise in response to various candidate aptamers.

Agency
National Institute of Health (NIH)
Institute
National Institute of Allergy and Infectious Diseases (NIAID)
Type
Research Project (R01)
Project #
5R01AI045344-02
Application #
6170339
Study Section
Special Emphasis Panel (ZRG1-AARR-1 (01))
Program Officer
Litterst, Charles L
Project Start
1999-07-15
Project End
2003-06-30
Budget Start
2000-07-01
Budget End
2001-06-30
Support Year
2
Fiscal Year
2000
Total Cost
$216,418
Indirect Cost
Name
Indiana University Bloomington
Department
Chemistry
Type
Schools of Arts and Sciences
DUNS #
006046700
City
Bloomington
State
IN
Country
United States
Zip Code
47401
Burke, Donald H; Greathouse, S Travis (2005) Low-magnesium, trans-cleavage activity by type III, tertiary stabilized hammerhead ribozymes with stem 1 discontinuities. BMC Biochem 6:14
Saksmerprome, Vanvimon; Roychowdhury-Saha, Manami; Jayasena, Sumedha et al. (2004) Artificial tertiary motifs stabilize trans-cleaving hammerhead ribozymes under conditions of submillimolar divalent ions and high temperatures. RNA 10:1916-24
Saksmerprome, Vanvimon; Burke, Donald H (2004) Deprotonation stimulates productive folding in allosteric TRAP hammerhead ribozymes. J Mol Biol 341:685-94
Saksmerprome, Vanvimon; Burke, Donald H (2003) Structural flexibility and the thermodynamics of helix exchange constrain attenuation and allosteric activation of hammerhead ribozyme TRAPs. Biochemistry 42:13879-86
Nickens, David G; Patterson, James T; Burke, Donald H (2003) Inhibition of HIV-1 reverse transcriptase by RNA aptamers in Escherichia coli. RNA 9:1029-33
Lato, Susan M; Ozerova, Nicole D S; He, Kaizhang et al. (2002) Boron-containing aptamers to ATP. Nucleic Acids Res 30:1401-7
Burke, Donald H; Nickens, David G (2002) Expressing RNA aptamers inside cells to reveal proteome and ribonome function. Brief Funct Genomic Proteomic 1:169-88
Burke, Donald H; Ozerova, Nicole D S; Nilsen-Hamilton, Marit (2002) Allosteric hammerhead ribozyme TRAPs. Biochemistry 41:6588-94