Isolation and Characterization of the Actin Gene from Tetrahymena thermophila

• Published in: Proceedings of the National Academy of Sciences - PNAS 1986-07, Vol.83 (14), p.5160-5164
• Main Authors: Cupples, Claire G., Pearlman, Ronald E.
• Format: Article
• Language: English
• Subjects: Actins; Amino acids; Biological and medical sciences; Biological Sciences: Genetics; Biotechnology; Cell motility; Codons; DNA; DNA probes; Fundamental and applied biological sciences. Psychology; Genes; Genetic engineering; Genetic technics; Methods. Procedures. Technologies; Molecular cloning; Sequencing; Stop codon; Yeasts
• ISSN: 0027-8424; 1091-6490
• DOI: 10.1073/pnas.83.14.5160

The macronucleus of Tetrahymena thermophila contains a single actin gene. We have isolated this gene from a partial plasmid library by using the yeast actin gene as a probe. The nucleotide sequence of the gene has been determined and the amino acid sequence of the potential protein deduced. The encoded protein is 375 amino acids long, one amino acid longer than the yeast actin. It is one of the most divergent actins sequenced to date, being only 75% homologous to yeast actin. Unlike the actin genes from most other organisms, it does not contain introns. The coding region contains TAA and TAG codons; the translation termination codon is TGA. Comparison of the amino acid sequence of the Tetrahymena actin with that of actins from other organisms suggests that TAG may code for glutamic acid. The gene is transcribed from multiple initiation sites between 57 and 98 nucleotides upstream of the translation start codon. The 5′ flanking region is very A+T-rich and contains numerous ``TATA-like'' sequences upstream of the transcription start sites.