Wheat acetyl-coenzyme A carboxylase: cDNA and protein structure

• Published in: Proceedings of the National Academy of Sciences - PNAS Vol. 91; no. 15; pp. 6860 - 6864
• Main Authors: Gornicki, P, Podkowinski, J, Scappino, L.A, DiMaio, J, Ward, E, Haselkorn, R
• Format: Journal Article
• Language: English
• Published: United States National Academy of Sciences of the United States of America 19.07.1994; National Acad Sciences; National Academy of Sciences
• Subjects: Acetyl-CoA Carboxylase - genetics; Amino Acid Sequence; Amino acids; Animals; ARN MENSAJERO; ARN MESSAGER; Base Sequence; Cellular biology; Chloroplasts; Complementary DNA; Deoxyribonucleic acid; Diatoms; DNA; Genetics; Humans; LIGASAS; LIGASE; Messenger RNA; Molecular Sequence Data; Plant cells; Polymerase Chain Reaction; Protein Conformation; Proteins; RNA; SECUENCIA NUCLEICA; Sequence Homology, Amino Acid; SEQUENCE NUCLEIQUE; Triticum - enzymology; Triticum - genetics; TRITICUM AESTIVUM; Wheat; Yeasts
• ISSN: 0027-8424; 1091-6490
• DOI: 10.1073/pnas.91.15.6860

cDNA fragments encoding part of wheat (Triticum aestivum) acetyl-CoA carboxylase (ACC; EC 6.4.1.2) were cloned by PCR using primers based on the alignment of several biotin-dependent carboxylases. A set of overlapping clones encoding the entire wheat ACC was then isolated by using these fragments as probes. The cDNA sequence contains a 2257-amino acid reading frame encoding a 251-kDa polypeptide. The amino acid sequence of the most highly conserved domain, corresponding to the biotin carboxylases of prokaryotes, is 52-55% identical to ACC of yeast, rat, and diatom. Identity with the available C-terminal amino acid sequence of maize ACC is 66%. The biotin attachment site has the typical eukaryotic EVMKM sequence. The cDNA does not encode an obvious chloroplast targeting sequence. Various cDNA fragments hybridize in Northern blots to a 7.9-kb mRNA. Southern analysis with cDNA probes revealed multiple hybridizing fragments in hexaploid wheat DNA. Some of the wheat cDNA probes also hybridize with ACC-specific DNA from other plants, indicating significant conservation among plant ACCs.