Polypeptide release factor eRF1 from Tetrahymena thermophila: cDNA cloning, purification and complex formation with yeast eRF3.
The first cDNA for the translational release factor eRF1 of ciliates was cloned from Tetrahymena thermophila. The coding frame contained one UAG and nine UAA codons that are reassigned for glutamine in Tetrahymena. The deduced protein sequence is 57% identical to human eRF1. The recombinant Tetrahymena eRF1 purified from a yeast expression system was able to bind to yeast eRF3 as do other yeast or mammalian eRF1s as a prerequisite step for protein termination. The recombinant Tetrahymena eRF1, nevertheless, failed to catalyze polypeptide termination in vitro with rat or Artemia ribosomes, at least in part, due to less efficient binding to the heterologous ribosomes. Stop codon specificity and phylogenetic significance of Tetrahymena eRF1 are discussed from the conservative protein feature.
Karamyshev
A L
AL
Department of Tumor Biology, Institute of Medical Science, University of Tokyo, 4-6-1 Shirokanedai, Minato-ku, Tokyo, Japan.
Ito
K
K
Nakamura
Y
Y
D000595:Amino Acid Sequence; D000818:Animals; D003001:Cloning, Molecular; D018076:DNA, Complementary / Q000032:analysis / Q000302:isolation & purification; D019281:Dimerization; D005656:Fungal Proteins / Q000378:metabolism*; D006801:Humans; D008969:Molecular Sequence Data; D010454:Peptide Termination Factors / Q000235:genetics* / Q000302:isolation & purification / Q000378:metabolism*; D010802:Phylogeny; D051381:Rats; D011994:Recombinant Proteins / Q000235:genetics / Q000302:isolation & purification / Q000378:metabolism; D012441:Saccharomyces cerevisiae / Q000166:cytology / Q000235:genetics / Q000378:metabolism; D012568:Schizosaccharomyces / Q000737:chemistry; D017386:Sequence Homology, Amino Acid; D016808:Tetrahymena thermophila / Q000737:chemistry / Q000235:genetics*
D018076:DNA, Complementary; C489002:ETF1 protein, human; C541746:Etf1 protein, rat; D005656:Fungal Proteins; D010454:Peptide Termination Factors; D011994:Recombinant Proteins; C087971:peptide-chain-release factor 3