Open AccessReplacement of anticodon loop nucleotides to produce functional tRNAs: amber suppressors derived from yeast tRNAPhe.
Author(s) -
A. Gregory Bruce,
John F. Atkins,
Norma M. Wills,
Olke C. Uhlenbeck,
Raymond F. Gesteland
Publication year - 1982
Publication title -
proceedings of the national academy of sciences of the united states of america
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 5.011
H-Index - 771
eISSN - 1091-6490
pISSN - 0027-8424
DOI - 10.1073/pnas.79.23.7127
Subject(s) - transfer rna , nucleotide , terminator (solar) , yeast , biology , genetic code , biochemistry , purine metabolism , protein biosynthesis , rna , genetics , amino acid , gene , enzyme , ionosphere , physics , astronomy
The method of anticodon loop replacement has been used to make derivatives of yeast tRNAPhe. By constructing tRNAs with a CUA anticodon, complementary to the amber (UAG) terminator, functional amber suppressor tRNAs were produced. The activity of these tRNAs was assayed in a mammalian cell-free protein synthesizing system. The level of suppression reflects the efficiency of codon recognition. tRNAs were constructed with either A, C, U, or G on the 3' side of the CUA anticodon. The tRNAs containing the purines were efficient amber suppressors, whereas those containing pyrimidines were inefficient.