Determination of the angle between the anticodon and aminoacyl acceptor stems of yeast phenylalanyl tRNA in solution.
A principal feature of the crystal structures of tRNAs is an L-shaped tertiary conformation in which the aminoacyl acceptor stem and the anticodon stem are approximately perpendicular. However, the anticodon-acceptor interstem angle has not been precisely quantified in solution for any tRNA. Such a determination would represent an important test of the predicted global conformation of tRNAs in solution. To this end, we have constructed a yeast tRNA(Phe) heteroduplex RNA molecule in which the anticodon and acceptor stems of the tRNA have each been extended by approximately 70 base pairs. A comparison of the rotational decay times of the heteroduplex molecule and a linear control yields an interstem angle of 89 +/- 4 degrees in 4 mM magnesium chloride/100 microM spermine hydrochloride, essentially identical to the corresponding angle observed in the crystal under similar buffer and temperature conditions. The current approach is applicable to the study of a wide variety of RNA molecules that possess elements of nonhelical structure.
Friederich
M W
MW
Department of Biochemistry, Biophysics, and Genetics, University of Colorado Health Sciences Center, Denver 80262, USA.
Gast
F U
FU
Vacano
E
E
Hagerman
P J
PJ
D000926:Anticodon / Q000737:chemistry* / Q000378:metabolism; D001483:Base Sequence; D007700:Kinetics; D015636:Magnesium Chloride / Q000494:pharmacology; D008958:Models, Molecular; D008969:Molecular Sequence Data; D009690:Nucleic Acid Conformation; D009692:Nucleic Acid Heteroduplexes / Q000737:chemistry / Q000378:metabolism; D010957:Plasmids; D012331:RNA, Fungal / Q000737:chemistry / Q000378:metabolism; D012360:RNA, Transfer, Phe / Q000737:chemistry* / Q000378:metabolism; D012441:Saccharomyces cerevisiae / Q000235:genetics / Q000378:metabolism*; D013997:Time Factors
D000926:Anticodon; D009692:Nucleic Acid Heteroduplexes; D012331:RNA, Fungal; D012360:RNA, Transfer, Phe; D015636:Magnesium Chloride