cDNA cloning and characterization of the nuclear gene encoding chloroplast glyceraldehyde-3-phosphate dehydrogenase from the marine red alga Gracilaria verrucosa

• Published in: Current genetics 1993-05, Vol.23 (5-6), p.483-489
• Main Authors: Zhou, Y.H. (National Research Council of Canada, Halifax, Nova Scotia (Canada). Inst. for Marine Biosciences), Ragan, M.A
• Format: Article
• Language: English
• Subjects: ADN; Amino Acid Sequence; Base Sequence; Biological and medical sciences; Blotting, Southern; cDNA; Cell Nucleus; Chloroplast isoenzyme; CHLOROPLASTE; Chloroplasts - enzymology; Cloning, Molecular; CLOROPLASTO; DNA; EVOLUCION; EVOLUTION; Fundamental and applied biological sciences. Psychology; GENE; GENES; Genes. Genome; Glyceraldehyde-3-phosphate dehydrogenase; Glyceraldehyde-3-Phosphate Dehydrogenases - genetics; Glyceraldehyde-3-Phosphate Dehydrogenases - metabolism; GRACILARIA; Gracilaria verrucosa; ISOENZIMAS; ISOENZYME; Molecular and cellular biology; Molecular evolution; Molecular genetics; Molecular Sequence Data; Polymerase Chain Reaction; Rhodophyta - enzymology; Rhodophyta - genetics; Sequence Homology, Amino Acid
• ISSN: 0172-8083; 1432-0983
• DOI: 10.1007/BF00312639

Using a PCR-generated homologous probe, cDNA was recovered (GapA cDNA) encoding the complete 338 amino-acid chloroplast GAPDH of the marine red alga Gracilaria verrucosa, together with its 78 amino- acid transit peptide. This cDNA was readily aligned with chloroplast-localized GAPDH genes (GapA and GapB) of green plants. The proline residue which contributes to the specificity of NAD(+) binding to cytosolic GAPDHs is absent from the deduced polypeptide chain of G. verrucosa GapA as is also the case in the chloroplast GAPDHs of plants. The transit peptide shows a high proportion of random coil, an amino-terminal Met-Ala dipeptide, a high content of hydroxylamino acids, and a net positive charge. The polyadenylation signal appears to be AGTAAA. Genomic Southern-hybridization data indicate that only one chloroplast-GAPDH gene may occur in G. verrucosa. Bootstrapped parsimony trees indicate that the G. verrucosa GapA gene is a sister group to plant chloroplast-GAPDH genes, and are most readily interpreted as showing that red algal and plant chloroplast-localized GAPDHs arose in a single endosymbiotic event.