Structural analyses on yeast tRNA(Tyr) and its complex with tyrosyl-tRNA synthetase by the use of hydroxyl radical 'footprinting'.
Hydroxyl radical, generated by reduction of hydrogen peroxide by Fe(II)-EDTA, was used to investigate the contact sites of yeast tRNA(Tyr) with its cognate tyrosyl-tRNA synthetase (TyrRS). Exposure of free tRNA(Tyr) to this reagent gave cleavage patterns consistent with the tertiary structure of yeast tRNA(Phe) established by X-ray crystallography. When the probing reaction was performed under the conditions which stabilized complex formation between tRNA(Tyr) and TyrRS, aminoacyl-stem region of the tRNA was protected from cleavage. This result supports our earlier finding that the information for binding to TyrRS would reside mainly in the aminoacyl-stem of tRNA(Tyr).
Motoki
I
I
Department of Biological Sciences, Faculty of Bioscience and Biotechnology, Tokyo Institute of Technology, Yokohama, Japan.
Yosinari
S
S
Watanabe
K
K
Nishikawa
K
K
D001665:Binding Sites; D004492:Edetic Acid; D005296:Ferrous Compounds; D005609:Free Radicals; D006861:Hydrogen Peroxide; D012331:RNA, Fungal / Q000737:chemistry / Q000378:metabolism; D012365:RNA, Transfer, Tyr / Q000737:chemistry* / Q000378:metabolism; D014448:Tyrosine-tRNA Ligase / Q000737:chemistry* / Q000378:metabolism
D005296:Ferrous Compounds; D005609:Free Radicals; D012331:RNA, Fungal; D012365:RNA, Transfer, Tyr; C055526:Fe(II)-EDTA; D004492:Edetic Acid; D006861:Hydrogen Peroxide; D014448:Tyrosine-tRNA Ligase