Design, biological activity and NMR-solution structure of a DNA analogue of yeast tRNA(Phe) anticodon domain.
Design of biologically active DNA analogues of the yeast tRNA(Phe) anticodon domain, tDNAPheAC, required the introduction of a d(m5C)-dependent, Mg(2+)-induced structural transition and the d(m1G) disruption of an intra-loop dC.dG base pair. The modifications were introduced at residues corresponding to m5C-40 and wybutosine-37 in tRNA(Phe). Modified tDNAPheAC inhibited translation by 50% at a tDNAPheAC:ribosome ratio of 8:1. The molecule's structure has been determined by NMR spectroscopy and restrained molecular dynamics with an overall r.m.s.d. of 2.8 A and 1.7 A in the stem, and is similar to the tRNA(Phe) anticodon domain in conformation and dimensions. The tDNAPheAC structure may provide a guide for the design of translation inhibitors as potential therapeutic agents.
Department of Biochemistry, North Carolina State University, Raleigh 27695-7622, USA.
M M
MM
Basti
J W
JW
Stuart
A T
AT
Lam
R
R
Guenther
P F
PF
Agris
D000926:Anticodon / Q000235:genetics*; D001483:Base Sequence; D004271:DNA, Fungal / Q000235:genetics* / Q000378:metabolism; D009682:Magnetic Resonance Spectroscopy; D008969:Molecular Sequence Data; D015394:Molecular Structure; D012331:RNA, Fungal / Q000235:genetics / Q000378:metabolism; D012360:RNA, Transfer, Phe / Q000235:genetics*; D012441:Saccharomyces cerevisiae / Q000235:genetics / Q000378:metabolism*
D000926:Anticodon; D004271:DNA, Fungal; D012331:RNA, Fungal; D012360:RNA, Transfer, Phe