The promoter sequence of a yeast tRNAtyr gene

Thirty-one base substitution mutations within the yeast SUP4 tRNAtyr gene were used to probe the effects of different intragenic sequences on promoter activity. The various mutant plasmids were tested quantitatively for their in vitro template activity and for their ability to block competitively the transcription of a reference gene. Five mutations within the coding sequence of SUP4 decreased template activity for pre-tRNAtyr synthesis. The competition assays revealed 11 mutant genes that behaved differently than SUP4-o. Six were weaker competitors and five were stronger. The 12 mutations affecting template activity or competition are clustered in three regions: those encoding the dihydrouracil (D) arm, the extra loop, and the T psi arm of the tRNA. All of the mutations that reduce competition involve base changes that decrease homology to a eucaryotic tRNA consensus sequence in the highly conserved D and T psi regions. Three of the five up mutations increased homology to the tRNA consensus sequence.