The major 5' determinant in stop codon read-through involves two adjacent adenines.
The aim of this approach was to identify the major determinants, located at the 5' end of the stop codon, that modulate translational read-through in Saccharomyces cerevisiae. We developed a library of oligonucleotides degenerate at the six positions immediately upstream of the termination codon, cloned in the ADE2 reporter gene. Variations at these positions modulated translational read-through efficiency approximately 16-fold. The major effect was imposed by the two nucleotides immediately upstream of the stop codon. We showed that this effect was neither mediated by the last amino acid residues present in the polypeptide chain nor by the tRNA present in the ribosomal P site. We propose that the mRNA structure, depending on the nucleotides in the P site, is the main 5' determinant of read-through efficiency.
Tork
Sanaa
S
CNRS UMR 8621, Institut de Génétique et Microbiologie, Université Paris-Sud, 91405 Orsay Cedex, France.
Hatin
Isabelle
I
Rousset
Jean-Pierre
JP
Fabret
Céline
C
D000227:Adenine Nucleotides / Q000235:genetics*; D001482:Base Composition; D001483:Base Sequence; D001665:Binding Sites; D018388:Codon, Terminator / Q000235:genetics*; D015723:Gene Library; D005800:Genes, Fungal / Q000235:genetics; D017930:Genes, Reporter / Q000235:genetics; D009690:Nucleic Acid Conformation; D009843:Oligoribonucleotides / Q000737:chemistry / Q000235:genetics / Q000378:metabolism; D010441:Peptide Chain Elongation, Translational / Q000235:genetics*; D012331:RNA, Fungal / Q000737:chemistry / Q000235:genetics / Q000378:metabolism; D012333:RNA, Messenger / Q000737:chemistry / Q000235:genetics / Q000378:metabolism; D012346:RNA, Transfer, Amino Acyl / Q000737:chemistry / Q000235:genetics / Q000378:metabolism; D038621:Regulatory Sequences, Ribonucleic Acid / Q000235:genetics*; D012270:Ribosomes / Q000235:genetics / Q000378:metabolism; D012441:Saccharomyces cerevisiae / Q000235:genetics*; D029701:Saccharomyces cerevisiae Proteins / Q000235:genetics
D000227:Adenine Nucleotides; D018388:Codon, Terminator; D009843:Oligoribonucleotides; D012331:RNA, Fungal; D012333:RNA, Messenger; D012346:RNA, Transfer, Amino Acyl; D038621:Regulatory Sequences, Ribonucleic Acid; D029701:Saccharomyces cerevisiae Proteins