This is a second revision of a proposal to measure certain structural features of tRNA in solution. Using a technique developed in his own laboratory, Dr. Hagerman can measure the angle between major stems in tRNA (and other RNAs) by comparing rotational diffusion times of a tRNA heteroduplex and a linear RNA of the same length. In particular, it is proposed to measure the """"""""L-angle"""""""" in tRNAs, the angle between the anti-codon and acceptor stems. For this work, the heteroduplex will be artificially extended by placing about 70 extra base pairs on the anti-codon and acceptor stems. Some mitochondrial tRNAs, for features suchas the D-loop or Tpsi C-loop appear to be missing. Recent modeling work suggests that these atypical tRNAs also possess atypical tRNA structure in 3 dimensions. Thus, the """"""""L-angle"""""""" for several of these atypical mitochondrial tRNAs will be measured to determine whether or not these models are consistent with data. The methods proposed here yield information not available from crystallographic or other solution studies. They can measure not only interstem angles, but also the range and flexibility of those angles. Thus the """"""""L-angle"""""""" and its range will be measured under different ionic conditions and with different tertiary interactions. Studying these gross structural changes when bases are altered is relevant to certain neuromuscular diseases caused by mutations in mitochondrial tRNAs.

Agency
National Institute of Health (NIH)
Institute
National Institute of General Medical Sciences (NIGMS)
Type
Research Project (R01)
Project #
5R01GM052557-03
Application #
2750010
Study Section
Special Emphasis Panel (ZRG3-BBCA (01))
Project Start
1996-08-01
Project End
2000-07-31
Budget Start
1998-08-01
Budget End
1999-07-31
Support Year
3
Fiscal Year
1998
Total Cost
Indirect Cost
Name
University of Colorado Denver
Department
Biochemistry
Type
Schools of Medicine
DUNS #
065391526
City
Aurora
State
CO
Country
United States
Zip Code
80045
Frazer-Abel, Ashley A; Hagerman, Paul J (2008) Core flexibility of a truncated metazoan mitochondrial tRNA. Nucleic Acids Res 36:5472-81
Tassone, F; Hagerman, R J; Taylor, A K et al. (2000) Clinical involvement and protein expression in individuals with the FMR1 premutation. Am J Med Genet 91:144-52
Frazer-Abel, A A; Hagerman, P J (1999) Determination of the angle between the acceptor and anticodon stems of a truncated mitochondrial tRNA. J Mol Biol 285:581-93
van Oppen, M J; Hislop, N R; Hagerman, P J et al. (1999) Gene content and organization in a segment of the mitochondrial genome of the scleractinian coral Acropora tenuis: major differences in gene order within the anthozoan subclass zoantharia. Mol Biol Evol 16:1812-5
Friederich, M W; Vacano, E; Hagerman, P J (1998) Global flexibility of tertiary structure in RNA: yeast tRNAPhe as a model system. Proc Natl Acad Sci U S A 95:3572-7
Friederich, M W; Hagerman, P J (1997) The angle between the anticodon and aminoacyl acceptor stems of yeast tRNA(Phe) is strongly modulated by magnesium ions. Biochemistry 36:6090-9