Abstract

The goal of this project is the determination of the structures of biologically relevant, small RNAs and RNA-protein complexes by NMR. It is anticipated that the data obtained will lead to a better understanding of the chemistry that underlies RNA function in normal gene expression, and increase the likelihood that therapeutic strategies can be developed based on the inhibition of RNA function. The systems to be studied include: (1) the alpha sarcin stem/loop from eucaryotic 28S rRNA, which plays a key role in the translocation step of protein synthesis, (2) pDG07 RNA, a deletion mutant of E. coli 5S rRNA that includes the binding site for ribosomal protein L25, (3) synthetic RNA that shows hammerhead activity, and (4) synthetic RNAs containing base pairing irregularities. The ribosomal protein L25/5S RNA complex from E. coli will also be analyzed. Since experiments that depend on labelling with stable isotopes will be required to solve these problems, strategies for labelling RNAs with isotopes will be pursued, and experimental approaches will be developed that make use of isotopically labelled molecules to increase the complexity of the RNA systems accessible to study by NMR.

  Funding Agency

Agency
National Institute of Health (NIH)
Institute
National Institute of General Medical Sciences (NIGMS)
Type
Research Project (R01)
Project #
5R01GM041651-06
Application #
2180981
Study Section
Molecular and Cellular Biophysics Study Section (BBCA)
Project Start
1989-07-01
Project End
1996-06-30
Budget Start
1994-07-01
Budget End
1995-06-30
Support Year
6
Fiscal Year
1994
Total Cost
Indirect Cost

  Institution

Name
Yale University
Department
Chemistry
Type
Schools of Arts and Sciences
DUNS #
082359691
City
New Haven
State
CT
Country
United States
Zip Code
06520

  Publications

Schweisguth, D C; Chelladurai, B S; Nicholson, A W et al. (1994) Structural characterization of a ribonuclease III processing signal. Nucleic Acids Res 22:604-12
Schweitzer, B I; Mikita, T; Kellogg, G W et al. (1994) Solution structure of a DNA dodecamer containing the anti-neoplastic agent arabinosylcytosine: combined use of NMR, restrained molecular dynamics, and full relaxation matrix refinement. Biochemistry 33:11460-75
Szewczak, A A; Moore, P B; Chang, Y L et al. (1993) The conformation of the sarcin/ricin loop from 28S ribosomal RNA. Proc Natl Acad Sci U S A 90:9581-5
Kellogg, G W; Schweitzer, B I (1993) Two- and three-dimensional 31P-driven NMR procedures for complete assignment of backbone resonances in oligodeoxyribonucleotides. J Biomol NMR 3:577-95
Szewczak, A A; Kellogg, G W; Moore, P B (1993) Assignment of NH resonances in nucleic acids using natural abundance 15N-1H correlation spectroscopy with spin-echo and gradient pulses. FEBS Lett 327:261-4
Cheong, C; Moore, P B (1992) Solution structure of an unusually stable RNA tetraplex containing G- and U-quartet structures. Biochemistry 31:8406-14
White, S A; Nilges, M; Huang, A et al. (1992) NMR analysis of helix I from the 5S RNA of Escherichia coli. Biochemistry 31:1610-21
Szewczak, A A; Chan, Y L; Moore, P B et al. (1991) On the conformation of the alpha sarcin stem-loop of 28S rRNA. Biochimie 73:871-7
Szewczak, A A; Webster, K R; Spicer, E K et al. (1991) An NMR characterization of the regA protein-binding site of bacteriophage T4 gene 44 mRNA. J Biol Chem 266:17832-7
Szewczak, A A; White, S A; Gewirth, D T et al. (1990) On the use of T7 RNA polymerase transcripts for physical investigation. Nucleic Acids Res 18:4139-42