Home LiteratureArticle Details
PMID: 7480350 Published · ppublish English Comparative Study Journal Article Research Support, Non-U.S. Gov't

Molecular cloning and characterization of the cDNA coding for the biotin-containing subunit of the chloroplastic acetyl-coenzyme A carboxylase.

Plant physiology ·Vol. 109 ·No. 2 ·1995-10-00 ·Pages 619-25

Choi JK, Yu F, Wurtele ES, Nikolau BJ

Abstract

We report the molecular cloning and sequence of the cDNA coding for the biotin-containing subunit of the chloroplastic acetylcoenzyme A (CoA) carboxylase (ACCase) of Arabidopsis thaliana (CAC1). The 3' end of the CAC1 sequence, coding for a peptide of 94 amino acids, which includes a putative biotinylation motif, was expressed in Escherichia coli as a glutathione-S-transferase (GST) fusion protein. The resulting GST-CAC1 fusion protein was biotinylated in vivo, indicating that CAC1 codes for a biotin-containing protein. Antibodies generated to the GST-CAC1 protein reacted solely with the 38-kD biotin-containing polypeptide of Arabidopsis. Furthermore, these antibodies inhibited ACCase activity in extracts from Arabidopsis leaves. The deduced amino acid sequence of CAC1 has an apparent N-terminal chloroplast-targeting transit peptide. The CAC1 protein is coded by a single Arabidopsis gene, and its mRNA accumulates to the highest levels in organs that are undergoing rapid growth. The amino acid sequence of the CAC1 protein is most similar to the biotin carboxyl-carrier protein component of eubacterial ACCases. These characterizations identify CAC1 as the biotin-containing subunit of the plastidic, heteromeric ACCase of Arabidopsis. The results support the ancient origin of the two structurally distinct ACCases of plants.

MeSH Terms
Acetyl-CoA Carboxylase/biosynthesis,chemistry,isolation & purification Amino Acid Sequence Arabidopsis/enzymology,genetics Bacteria/enzymology Base Sequence Biotin Chloroplasts/enzymology Cloning, Molecular DNA, Complementary Macromolecular Substances Molecular Sequence Data Recombinant Proteins/biosynthesis,chemistry,isolation & purification Sequence Homology, Amino Acid
Chemicals
DNA, Complementary Macromolecular Substances Recombinant Proteins Biotin Acetyl-CoA Carboxylase
Authors & Affiliations
4 authors, click to expand affiliations / ORCID
Choi J K
Department of Biochemistry and Biophysics, Iowa State University, Ames 50011, USA.
Yu F
Wurtele E S
Nikolau B J
References (42)
42 references, click to expand
  1. Single-step purification of polypeptides expressed in Escherichia coli as fusions with glutathione S-transferase.
    Gene. 1988 Jul 15;67(1):31-40 PMID: 3047011
  2. Isolation of cDNA clones coding for the alpha and beta chains of human propionyl-CoA carboxylase: chromosomal assignments and DNA polymorphisms associated with PCCA and PCCB genes.
    Proc Natl Acad Sci U S A. 1986 Jul;83(13):4864-8 PMID: 3460076
  3. Amino acid sequence of the biotinyl subunit from transcarboxylase.
    J Biol Chem. 1979 Nov 25;254(22):11615-22 PMID: 40985
  4. Structural analysis, plastid localization, and expression of the biotin carboxylase subunit of acetyl-coenzyme A carboxylase from tobacco.
    Plant Physiol. 1995 Jun;108(2):805-12 PMID: 7610168
  5. Acetyl-CoA carboxylase from yeast is an essential enzyme and is regulated by factors that control phospholipid metabolism.
    J Biol Chem. 1993 May 25;268(15):10946-52 PMID: 8098706
  6. Chloroplast-encoded protein as a subunit of acetyl-CoA carboxylase in pea plant.
    J Biol Chem. 1993 Nov 25;268(33):25118-23 PMID: 7901221
  7. Sequence and transcriptional analysis of the gene cluster trnQ-zfpA-psaI-ORF231-petA in pea chloroplasts.
    Curr Genet. 1991 Nov;20(5):431-6 PMID: 1807835
  8. Structure and expression of an Arabidopsis acetyl-coenzyme A carboxylase gene.
    Plant Physiol. 1994 Jun;105(2):611-7 PMID: 7915036
  9. Molecular cloning and characterization of the cDNA coding for the biotin-containing subunit of 3-methylcrotonoyl-CoA carboxylase: identification of the biotin carboxylase and biotin-carrier domains.
    Proc Natl Acad Sci U S A. 1994 Jun 21;91(13):5779-83 PMID: 8016064
  10. Saccharopolyspora hirsuta 367 encodes clustered genes similar to ketoacyl synthase, ketoacyl reductase, acyl carrier protein, and biotin carboxyl carrier protein.
    Mol Gen Genet. 1993 Jul;240(1):146-50 PMID: 8341260
  11. Molecular cloning, characterization, and elicitation of acetyl-CoA carboxylase from alfalfa.
    Proc Natl Acad Sci U S A. 1994 May 10;91(10):4323-7 PMID: 7910406
  12. Localization and characterization of two structurally different forms of acetyl-CoA carboxylase in young pea leaves, of which one is sensitive to aryloxyphenoxypropionate herbicides.
    Biochem J. 1994 Jun 1;300 ( Pt 2):557-65 PMID: 7911659
  13. Fat metabolism in higher plants. LIV. A procaryotic type acetyl CoA carboxylase in spinach chloroplasts.
    Arch Biochem Biophys. 1972 Sep;152(1):83-91 PMID: 4561257
  14. Use of streptavidin to detect biotin-containing proteins in plants.
    Anal Biochem. 1985 Sep;149(2):448-53 PMID: 2866733
  15. Size determination of multienzyme complexes using two-dimensional agarose gel electrophoresis.
    Proteins. 1989;5(3):224-32 PMID: 2506543
  16. Cloning of the yeast FAS3 gene and primary structure of yeast acetyl-CoA carboxylase.
    Proc Natl Acad Sci U S A. 1992 May 15;89(10):4534-8 PMID: 1350093
  17. Characterization of Maize Acetyl-Coenzyme A Carboxylase.
    Plant Physiol. 1993 Feb;101(2):499-506 PMID: 12231704
  18. Improved purification and further characterization of acetyl-CoA carboxylase from cultured cells of parsley (Petroselinum hortense).
    Eur J Biochem. 1983 Jun 15;133(2):335-9 PMID: 6133748
  19. Plants contain multiple biotin enzymes: discovery of 3-methylcrotonyl-CoA carboxylase, propionyl-CoA carboxylase and pyruvate carboxylase in the plant kingdom.
    Arch Biochem Biophys. 1990 Apr;278(1):179-86 PMID: 2321957
  20. Subcellular localization of acyl carrier protein in leaf protoplasts of Spinacia oleracea.
    Proc Natl Acad Sci U S A. 1979 Mar;76(3):1194-8 PMID: 286305
  21. The genes encoding the two carboxyltransferase subunits of Escherichia coli acetyl-CoA carboxylase.
    J Biol Chem. 1992 Aug 25;267(24):16841-7 PMID: 1355089
  22. Biosynthesis of C(20) and C(22) Fatty Acids by Developing Seeds of Limnanthes alba: CHAIN ELONGATION AND Delta5 DESATURATION.
    Plant Physiol. 1980 Oct;66(4):649-55 PMID: 16661496
  23. Evolutionary conservation among biotin enzymes.
    J Biol Chem. 1988 May 15;263(14):6461-4 PMID: 2896195
  24. Fatty acid synthesis and its regulation.
    Annu Rev Biochem. 1983;52:537-79 PMID: 6137188
  25. Cloning and characterization of the gene that encodes acetyl-coenzyme A carboxylase in the alga Cyclotella cryptica.
    J Biol Chem. 1993 Sep 15;268(26):19254-9 PMID: 8103514
  26. Molecular cloning of cDNAs and genes coding for beta-methylcrotonyl-CoA carboxylase of tomato.
    J Biol Chem. 1994 Apr 22;269(16):11760-8 PMID: 8163472
  27. The sodium ion translocating oxalacetate decarboxylase of Klebsiella pneumoniae. Sequence of the biotin-containing alpha-subunit and relationship to other biotin-containing enzymes.
    J Biol Chem. 1988 Jul 15;263(20):9640-5 PMID: 2454915
  28. The gene encoding the biotin carboxylase subunit of Escherichia coli acetyl-CoA carboxylase.
    J Biol Chem. 1992 Jan 15;267(2):855-63 PMID: 1370469
  29. Acyl-coenzyme A carboxylase of the free-living nematode Turbatrix aceti. 1. Its isolation and molecular characteristics.
    Biochemistry. 1978 May 16;17(10):1822-7 PMID: 656363
  30. Organization and nucleotide sequences of the genes encoding the biotin carboxyl carrier protein and biotin carboxylase protein of Pseudomonas aeruginosa acetyl coenzyme A carboxylase.
    J Bacteriol. 1993 Nov;175(21):6881-9 PMID: 7693652
  31. Acetyl coenzyme A carboxylase system of Escherichia coli. Purification and properties of the biotin carboxylase, carboxyltransferase, and carboxyl carrier protein components.
    J Biol Chem. 1974 Oct 25;249(20):6633-45 PMID: 4154089
  32. Structure of the coding sequence and primary amino acid sequence of acetyl-coenzyme A carboxylase.
    Proc Natl Acad Sci U S A. 1988 Aug;85(16):5784-8 PMID: 2901088
  33. Acetyl-CoA carboxylase from Escherichia coli: gene organization and nucleotide sequence of the biotin carboxylase subunit.
    Proc Natl Acad Sci U S A. 1991 Nov 1;88(21):9730-3 PMID: 1682920
  34. Genes for two subunits of acetyl coenzyme A carboxylase of Anabaena sp. strain PCC 7120: biotin carboxylase and biotin carboxyl carrier protein.
    J Bacteriol. 1993 Aug;175(16):5268-72 PMID: 8102363
  35. A gene encoding acetyl-coenzyme A carboxylase from Brassica napus.
    Plant Physiol. 1994 Oct;106(2):793-4 PMID: 7991694
  36. Acetyl coenzyme A carboxylase.
    Curr Top Cell Regul. 1974;8(0):139-95 PMID: 4154092
  37. Compartmentalization of two forms of acetyl-CoA carboxylase in plants and the origin of their tolerance toward herbicides.
    Proc Natl Acad Sci U S A. 1994 Apr 26;91(9):3598-601 PMID: 7909603
  38. Conformational flexibility and folding of synthetic peptides representing an interdomain segment of polypeptide chain in the pyruvate dehydrogenase multienzyme complex of Escherichia coli.
    J Biol Chem. 1989 Jan 15;264(2):767-75 PMID: 2642904
  39. Primary structure of chicken liver acetyl-CoA carboxylase deduced from cDNA sequence.
    J Biol Chem. 1988 Feb 25;263(6):2651-7 PMID: 2893793
  40. Wheat acetyl-coenzyme A carboxylase: cDNA and protein structure.
    Proc Natl Acad Sci U S A. 1994 Jul 19;91(15):6860-4 PMID: 7913745
  41. Molecular cloning of the biotinylated subunit of 3-methylcrotonyl-coenzyme A carboxylase of Arabidopsis thaliana.
    Plant Physiol. 1995 Mar;107(3):1013-4 PMID: 7716229
  42. Adipose pyruvate carboxylase: amino acid sequence and domain structure deduced from cDNA sequencing.
    Proc Natl Acad Sci U S A. 1993 Mar 1;90(5):1766-70 PMID: 8446588
Article Info
Journal
Plant physiology
Abbr.
Plant Physiol
ISSN
0032-0889
DOI
10.1104/pp.109.2.619
Published
1995-10-00
Pages
619-25
Language
English
Region
United States
NLM ID
0401224
PMCID
PMC157628
Subset
IM
Databases
GENBANK
U23155
External Links
PubMed source DOI: 10.1104/pp.109.2.619 PMC Full Text View journal details
Analysis Services
Analysis Services

Contact

No. 2 Wenbo Road, Zhangqiu District, Jinan, Shandong

Qilu Normal University · Genelibs Bioinformatics Lab

750 Shunhua Rd, Jinan

2F, Bldg F, University Science Park

Tel: 0531-88819269

WeChat Official Account

Follow our WeChat subscription account for real-time updates and the latest in medical and biological research.


Business Email

E-mail: [email protected]

Copyright © 2026.Genelibs Jinan Honghui Biotechnology Co., Ltd. 鲁ICP备20007159号-2