Isolation and Structural Characterization of cDNA Clones Encoding the Mating Pheromone Er-1 Secreted by the Ciliate Euplotes raikovi

• Published in: Proceedings of the National Academy of Sciences - PNAS Vol. 86; no. 9; pp. 3016 - 3020
• Main Authors: Miceli, Cristina, La Terza, Antonietta, Melli, Marialuisa
• Format: Journal Article
• Language: English
• Published: Washington, DC National Academy of Sciences of the United States of America 01-05-1989; National Acad Sciences
• Subjects: Amino Acid Sequence; Amino acids; Animals; Base Sequence; Biological and medical sciences; cDNA; Ciliophora - genetics; Cloning, Molecular; Codon; Codons; Complementary DNA; DNA; DNA - genetics; DNA - isolation & purification; Euplotes raikovi; Fundamental and applied biological sciences. Psychology; Gels; Genes. Genome; Membrane Proteins; Molecular and cellular biology; Molecular genetics; Molecular Sequence Data; Nucleic Acid Hybridization; nucleotide sequence; Oligonucleotide Probes; Oligonucleotides; Open reading frames; Peptides - genetics; Pheromones; Poly A; Protein Biosynthesis; Protozoan Proteins; RNA; RNA - genetics; RNA, Messenger - genetics; Sequencing; Transcription, Genetic; Characterization; C terminal-Sequence; Complementary DNA; Blotting assay; Isolation; Pheromone; Genetic engineering; Molecular probe; Inverted repeated sequence; Aminoacid sequence
• ISSN: 0027-8424; 1091-6490
• DOI: 10.1073/pnas.86.9.3016

cDNA clones comprising the entire coding region for the mating pheromone Er-1 of Euplotes raikovi have been isolated by oligonucleotide screening of two cDNA libraries in the vectors λ gt10 and pUC12. The cDNA sequence contains an open reading frame of 75 amino acids that constitute pre-pro-Er-1. The amino acid sequence of secreted Er-1 starts at aspartic acid-36 of pre-pro-Er-1 and completely matches that known by direct Er-1 protein sequencing. The coding region of Er-1 cDNA ends with codon TAA, which specifies glutamine in other ciliates. The 5′- and 3′-noncoding regions contain, respectively, two and one inverted repeats. The 3′-noncoding-region inverted repeat, which includes the unusual polyadenylylation signal AACAAA, has been related to RNA 3′-terminus formation.