Characterization and cloning of glycogen phosphorylase 1 from Dictyostelium discoideum

• Published in: Biochimica et biophysica acta Vol. 1129; no. 3; pp. 262 - 272
• Main Authors: Rogers, Patricia V., Luo, Shun, Sucic, Joseph F., Rutherford, Charles L.
• Format: Journal Article
• Language: English
• Published: Amsterdam Elsevier B.V 11.02.1992; Elsevier
• Subjects: Amino Acid Sequence; Animals; Base Sequence; Biological and medical sciences; Cloning, Molecular; Codon usage; Developmental mRNA expression; Dictyostelium - enzymology; Dictyostelium - growth & development; DNA - genetics; Fundamental and applied biological sciences. Psychology; Genes. Genome; Genomic copy number; Immunoblotting; Intron; Inverted PCR; Molecular and cellular biology; Molecular genetics; Molecular Sequence Data; Phosphorylases - genetics; Polymerase Chain Reaction; Restriction Mapping; RNA, Fungal - genetics; Secondary and tertiary protein structure; Sequence Homology, Nucleic Acid; Secondary and tertiary protein structure; ORF; Genomic copy number; Intron; AMP; Developmental mRNA expression; Codon usage; gp2; gp1; GCG; Inverted PCR; PCR; Dictyostelium discoideum; Polymerase chain reaction; Protozoa; Nucleotide sequence; Enzyme; Glycogen phosphorylase; Primary structure; Molecular cloning; Gene expression; Comparative study; Aminoacid sequence; Mycetozoa
• ISSN: 0167-4781; 0006-3002; 1879-2634
• DOI: 10.1016/0167-4781(92)90502-Q

We have cloned cDNA and genomic DNA fragments from Dictyostelium discoiseum that represent the entire coding egion of glycogen phosphorylase 1 (gpl, α- d-glucosyltransferase, EC 2.4.1.1). Nucleic acid sequencing of the gp1 clones revealed a single 139 bp intron separating the two exons that encode the 853 amino acids of gp1. The gp1 sequences are similar to other genes and proteins described for Dictyostelium in terms of G + C composition of coding and noncoding regions, splice junctions, intron length, codon preference and termination signals. Genomic Southern blot hybridizations suggest that gp1 exists as a single or low copy number gene in Dictyostelium. Northern analyses demonstrate that gp1 is a developmentally regulated transcript. In alignments of the gp1 peptide sequence to glycogen phosphorylase sequences from other organisms, a high degree of amino acid conservation at many active and regulatory sites was found; however, critical residues in the AMP and purine binding sites were not conserved.