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. 1996 Apr;110(4):1197–1205. doi: 10.1104/pp.110.4.1197

Characterization of soybean choline kinase cDNAs and their expression in yeast and Escherichia coli.

D E Monks 1, J H Goode 1, R E Dewey 1
PMCID: PMC160908  PMID: 8934624

Abstract

An expressed sequence tag from Arabidopsis that displayed sequence homology to mammalian and yeast choline kinases was used to isolate choline kinase-like cDNAs from soybean (Glycine max L.). Two distinct cDNAs, designated GmCK1 and GmCK2, were recovered that possessed full-length reading frames, each sharing approximately 32% identity at the predicted amino acid level with the rat choline kinase. A third unique choline kinase-like cDNA, GmCK3, was also identified but was not full length. Heterologous expression of GmCK1 in yeast (Saccharomyces cerevisiae) and GmCK2 in both yeast and Escherichia coli demonstrated that each encodes choline kinase activity. In addition to choline, other potential substrates for the choline kinase enzyme include ethanolamine, monomethylethanolamine (MME), and dimethylethanolamine (DME). Both soybean choline kinase isoforms demonstrated negligible ethanolamine kinase activity. Competitive inhibition assays, however, revealed very distinct differences in their responses to DME and MME. DME effectively inhibited only the GmCK2-encoded choline kinase activity. Although MME failed to effectively inhibit either reaction, an unexpected enhancement of choline kinase activity was observed specifically with the GmCK1-encoded enzyme. These results show that choline kinase is encoded by a small, multigene family in soybean comprising two or more distinct isoforms that exhibit both similarities and differences with regard to substrate specificity.

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Selected References

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  1. Becker D. M., Fikes J. D., Guarente L. A cDNA encoding a human CCAAT-binding protein cloned by functional complementation in yeast. Proc Natl Acad Sci U S A. 1991 Mar 1;88(5):1968–1972. doi: 10.1073/pnas.88.5.1968. [DOI] [PMC free article] [PubMed] [Google Scholar]
  2. Carman G. M., Henry S. A. Phospholipid biosynthesis in yeast. Annu Rev Biochem. 1989;58:635–669. doi: 10.1146/annurev.bi.58.070189.003223. [DOI] [PubMed] [Google Scholar]
  3. Datko A. H., Mudd S. H. Enzymes of phosphatidylcholine synthesis in lemna, soybean, and carrot. Plant Physiol. 1988 Dec;88(4):1338–1348. doi: 10.1104/pp.88.4.1338. [DOI] [PMC free article] [PubMed] [Google Scholar]
  4. Datko A. H., Mudd S. H. Phosphatidylcholine synthesis: differing patterns in soybean and carrot. Plant Physiol. 1988 Nov;88(3):854–861. doi: 10.1104/pp.88.3.854. [DOI] [PMC free article] [PubMed] [Google Scholar]
  5. Dewey R. E., Wilson R. F., Novitzky W. P., Goode J. H. The AAPT1 gene of soybean complements a cholinephosphotransferase-deficient mutant of yeast. Plant Cell. 1994 Oct;6(10):1495–1507. doi: 10.1105/tpc.6.10.1495. [DOI] [PMC free article] [PubMed] [Google Scholar]
  6. Feinberg A. P., Vogelstein B. A technique for radiolabeling DNA restriction endonuclease fragments to high specific activity. Anal Biochem. 1983 Jul 1;132(1):6–13. doi: 10.1016/0003-2697(83)90418-9. [DOI] [PubMed] [Google Scholar]
  7. Gietz D., St Jean A., Woods R. A., Schiestl R. H. Improved method for high efficiency transformation of intact yeast cells. Nucleic Acids Res. 1992 Mar 25;20(6):1425–1425. doi: 10.1093/nar/20.6.1425. [DOI] [PMC free article] [PubMed] [Google Scholar]
  8. Hosaka K., Kodaki T., Yamashita S. Cloning and characterization of the yeast CKI gene encoding choline kinase and its expression in Escherichia coli. J Biol Chem. 1989 Feb 5;264(4):2053–2059. [PubMed] [Google Scholar]
  9. Hosaka K., Tanaka S., Nikawa J., Yamashita S. Cloning of a human choline kinase cDNA by complementation of the yeast cki mutation. FEBS Lett. 1992 Jun 15;304(2-3):229–232. doi: 10.1016/0014-5793(92)80625-q. [DOI] [PubMed] [Google Scholar]
  10. Kinney A. J., Moore T. S., Jr Phosphatidylcholine synthesis in castor bean endosperm: characteristics and reversibility of the choline kinase reaction. Arch Biochem Biophys. 1988 Jan;260(1):102–108. doi: 10.1016/0003-9861(88)90429-8. [DOI] [PubMed] [Google Scholar]
  11. McGee T. P., Skinner H. B., Bankaitis V. A. Functional redundancy of CDP-ethanolamine and CDP-choline pathway enzymes in phospholipid biosynthesis: ethanolamine-dependent effects on steady-state membrane phospholipid composition in Saccharomyces cerevisiae. J Bacteriol. 1994 Nov;176(22):6861–6868. doi: 10.1128/jb.176.22.6861-6868.1994. [DOI] [PMC free article] [PubMed] [Google Scholar]
  12. Mellor R. B., Christensen T. M., Werner D. Choline kinase II is present only in nodules that synthesize stable peribacteroid membranes. Proc Natl Acad Sci U S A. 1986 Feb;83(3):659–663. doi: 10.1073/pnas.83.3.659. [DOI] [PMC free article] [PubMed] [Google Scholar]
  13. Messing J. Cloning in M13 phage or how to use biology at its best. Gene. 1991 Apr;100:3–12. doi: 10.1016/0378-1119(91)90344-b. [DOI] [PubMed] [Google Scholar]
  14. Mudd S. H., Datko A. H. Phosphoethanolamine bases as intermediates in phosphatidylcholine synthesis by lemna. Plant Physiol. 1986 Sep;82(1):126–135. doi: 10.1104/pp.82.1.126. [DOI] [PMC free article] [PubMed] [Google Scholar]
  15. Mudd S. H., Datko A. H. Synthesis of methylated ethanolamine moieties: regulation by choline in soybean and carrot. Plant Physiol. 1989 May;90(1):306–310. doi: 10.1104/pp.90.1.306. [DOI] [PMC free article] [PubMed] [Google Scholar]
  16. Newman T., de Bruijn F. J., Green P., Keegstra K., Kende H., McIntosh L., Ohlrogge J., Raikhel N., Somerville S., Thomashow M. Genes galore: a summary of methods for accessing results from large-scale partial sequencing of anonymous Arabidopsis cDNA clones. Plant Physiol. 1994 Dec;106(4):1241–1255. doi: 10.1104/pp.106.4.1241. [DOI] [PMC free article] [PubMed] [Google Scholar]
  17. Prud'homme M. P., Moore T. S. Phosphatidylcholine synthesis in castor bean endosperm : free bases as intermediates. Plant Physiol. 1992 Nov;100(3):1527–1535. doi: 10.1104/pp.100.3.1527. [DOI] [PMC free article] [PubMed] [Google Scholar]
  18. RAMASARMA T., WETTER L. R. Choline kinase of rapeseed (Brassica campestris L.). Can J Biochem Physiol. 1957 Oct;35(10):853–863. [PubMed] [Google Scholar]
  19. Tanaka K., Tolbert N. E., Gohlke A. F. Choline kinase and phosphorylcholine phosphatase in plants. Plant Physiol. 1966 Feb;41(2):307–312. doi: 10.1104/pp.41.2.307. [DOI] [PMC free article] [PubMed] [Google Scholar]
  20. Uchida T., Yamashita S. Molecular cloning, characterization, and expression in Escherichia coli of a cDNA encoding mammalian choline kinase. J Biol Chem. 1992 May 15;267(14):10156–10162. [PubMed] [Google Scholar]
  21. Uchida T., Yamashita S. Purification and properties of choline kinase from rat brain. Biochim Biophys Acta. 1990 Apr 17;1043(3):281–288. doi: 10.1016/0005-2760(90)90028-v. [DOI] [PubMed] [Google Scholar]
  22. WITTENBERG J., KORNBERG A. Choline phosphokinase. J Biol Chem. 1953 May;202(1):431–444. [PubMed] [Google Scholar]

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