Cloning and cDNA sequence of the regulatory subunit of cAMP-dependent protein kinase from Dictyostelium discoideum

• Published in: Proceedings of the National Academy of Sciences - PNAS Vol. 84; no. 1; pp. 6 - 10
• Main Authors: Mutzel, R, Lacombe, M.L, Simon, M.N, Gunzburg, J. de, Veron, M
• Format: Journal Article
• Language: English
• Published: Washington, DC National Academy of Sciences of the United States of America 01-01-1987; National Acad Sciences
• Subjects: ADN; Amino Acid Sequence; Amino acids; Animals; Base Sequence; Binding sites; Biochemistry; Biological and medical sciences; Biotechnology; Carrier Proteins - genetics; Cattle; cDNA; CHAMPIGNON; CLONE; CLONES; Cloning, Molecular; Complementary DNA; Dictyostelium - enzymology; Dictyostelium - genetics; Dictyostelium discoideum; DNA; DNA - metabolism; Eukaryotic cells; Fundamental and applied biological sciences. Psychology; FUNGI; Generally accepted auditing standards; Genes; Genes, Fungal; Genetic engineering; Genetic technics; Intracellular Signaling Peptides and Proteins; Methods. Procedures. Technologies; Molecular cloning; Myocardium - enzymology; NUCLEOTIDE; nucleotide sequence; Nucleotide sequences; NUCLEOTIDES; NUCLEOTIDOS; Open reading frames; protein kinase; Protein Kinases - genetics; Sequence Homology, Nucleic Acid; TRANSFERASAS; TRANSFERASE; TRANSFERASES; Ungulates; Yeasts; Dictyostelium discoideum; Protozoa; Complementary DNA; Protein kinase; Biological evolution; Primary structure; Models; Regulatory subunit; Molecular cloning; Mycetozoa
• ISSN: 0027-8424; 1091-6490
• DOI: 10.1073/pnas.84.1.6

cDNA clones encoding the regulatory subunit of the cAMP-dependent protein kinase (ATP:protein phosphotransferase, EC 2.7.1.37) from Dictyostelium discoideum were isolated by immunoscreening of a cDNA library constructed in the expression vector λ gt11. High-affinity cAMP-binding activity was detected in extracts from bacteria lysogenized with these clones. Nucleotide sequence analysis of three overlapping clones allowed the determination of a 1195-base-pair cDNA sequence coding for the entire regulatory subunit and containing nontranslated 5′ and 3′ sequences. The open reading frame codes for a protein of 327 amino acids, with molecular weight 36,794. The regulatory subunit from Dictyostelium shares a high degree of homology with its mammalian counterparts, but is lacking the NH2-terminal domain required for the association of regulatory subunits into dimers in other eukaryotes. On the basis of the comparison of the regulatory subunits from Dictyostelium, yeast, and bovine tissues, a model for the evolution of these proteins is proposed.