Fig. 2.

Transplantation of editing domains into iodoTyrRS. (A) The domains of iodoTyrRS and its variants. Numbers indicate the first residue of each domain. A filled symbol represents each enzyme, and the same symbol is used to indicate the data for the enzyme in B–G. (B and D) Time courses of tyrosyl-tRNA^Tyr formation in the presence of iodoTyrRS, the indicated fusion/insertion variants, the indicated editing-deficient fusion variants, and no enzyme. (C and E) Formation of tyrosyl-tRNA^Tyr (lanes Y) and iodotyrosyl-tRNA^Tyr (lanes IY) in the presence of the indicated enzymes. The bands corresponding to aminoacyl-tRNA^Tyr and uncharged tRNA^Tyr are marked. (F) Time courses of tyrosyl-tRNA^Tyr hydrolysis in the presence of N-Ped-IYRS, CP-Ped-IYRS, and no enzyme. (G) The translation of GST(Am) in the wheat germ cell-free system. The indicated enzymes were added to the reactions, together with the suppressor tRNA, in the absence (−IY) and presence (IY) of 3-iodo-l-tyrosine. All of the reactions contained l-tyrosine. The produced full-length GST was detected by Western blotting.