Effects of modification of 4-thiouridine in E. coli tRNA(fMet) on its methyl acceptor activity by thermostable Gm-methylases.

Journal of Biochemistry
H HoriK Watanabe

Abstract

tRNA(guanosine-2'-)-methyltransferases (Gm-methylases) isolated from extreme thermophiles, Thermus thermophilus strains HB 27 and HB 8, methylate the 2'-OH of the G18 ribose of the GG sequence in the D loop of tRNA, by recognizing the D "loop-stem" structure as a minimal requirement. To examine the role of the consensus uridine residue at position 8 (U8) adjacent to the D "loop-stem" region in the recognition of Gm-methylase, 4-thiouridine at this position (s4U8) in Escherichia coli tRNAfMet was modified reversibly with S-benzylthioisothiourea (sBTIU) or irreversibly by UV light. The initial velocities of the methylation reaction for the sBTIU-modified and the UV-induced cross-linked tRNAs were decreased to 40 and 30%, respectively, of that of the intact tRNA, but the sBTIU-modified tRNA regained almost full activity on reduction with beta-mercaptoethanol. Although both of the modified tRNAfMetS showed larger Km (although to different extents) and slightly smaller Vmax than the intact tRNAfMet, they retained full activities of methylation with tRNA(adenine-1-)-methyltransferase (m1A-methylase) and of aminoacylation with aminoacyl-tRNA synthetase (ARS) fraction as well, both of which were prepared from T. thermophilus strain HB ...Continue Reading

Citations

Mar 29, 2002·Genes to Cells : Devoted to Molecular & Cellular Mechanisms·Hiroyuki HoriKimitsuna Watanabe
Nov 24, 2006·Genes to Cells : Devoted to Molecular & Cellular Mechanisms·Hiroshi TakedaHiroyuki Hori
Jan 8, 2005·The Journal of Biological Chemistry·Kazunori WatanabeHiroyuki Hori
Sep 25, 1998·The Journal of Biological Chemistry·H HoriK Watanabe
May 1, 2014·EcoSal Plus·Glenn R Björk, Tord G Hagervall
Nov 1, 2005·EcoSal Plus·Glenn R Björk, Tord G Hagervall

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