Skip to main content
Journal of Bacteriology logoLink to Journal of Bacteriology
. 1988 Jun;170(6):2763–2769. doi: 10.1128/jb.170.6.2763-2769.1988

Nucleotide sequence of aceK, the gene encoding isocitrate dehydrogenase kinase/phosphatase.

D J Klumpp 1, D W Plank 1, L J Bowdin 1, C S Stueland 1, T Chung 1, D C LaPorte 1
PMCID: PMC211200  PMID: 2836370

Abstract

In Escherichia coli, the phosphorylation and dephosphorylation of isocitrate dehydrogenase (IDH) are catalyzed by a bifunctional protein kinase/phosphatase. We have determined the nucleotide sequence of aceK, the gene encoding IDH kinase/phosphatase. This gene consists of a single open reading frame of 1,734 base pairs preceded by a Shine-Dalgarno ribosome-binding site. Examination of the deduced amino acid sequence of IDH kinase/phosphatase revealed sequences which are similar to the consensus sequence for ATP-binding sites. This protein did not, however, exhibit the extensive sequence homologies which are typical of other protein kinases. Multiple copies of the REP family of repetitive extragenic elements were found within the intergenic region between aceA (encoding isocitrate lyase) and aceK. These elements have the potential for combining to form an exceptionally stable stem-loop structure (delta G = -54 kcal/mol [ca. -226 kJ/mol]) in the mRNA. This structure, which masks the ribosome-binding site and start codon for aceK, may contribute to the downshift in expression observed between aceA and aceK. Another potential stem-loop structure (delta G = -29 kcal/mol [ca. 121 kJ/mol]), unrelated to the REP sequences, was found within aceK.

Full text

PDF
2763

Images in this article

Selected References

These references are in PubMed. This may not be the complete list of references from this article.

  1. Belfaiza J., Parsot C., Martel A., de la Tour C. B., Margarita D., Cohen G. N., Saint-Girons I. Evolution in biosynthetic pathways: two enzymes catalyzing consecutive steps in methionine biosynthesis originate from a common ancestor and possess a similar regulatory region. Proc Natl Acad Sci U S A. 1986 Feb;83(4):867–871. doi: 10.1073/pnas.83.4.867. [DOI] [PMC free article] [PubMed] [Google Scholar]
  2. Bennett P. M., Holms W. H. Reversible inactivation of the isocitrate dehydrogenase of Escherichia coli ML308 during growth on acetate. J Gen Microbiol. 1975 Mar;87(1):37–51. doi: 10.1099/00221287-87-1-37. [DOI] [PubMed] [Google Scholar]
  3. Borthwick A. C., Holms W. H., Nimmo H. G. Amino acid sequence round the site of phosphorylation in isocitrate dehydrogenase from Escherichia coli ML308. FEBS Lett. 1984 Aug 20;174(1):112–115. doi: 10.1016/0014-5793(84)81087-x. [DOI] [PubMed] [Google Scholar]
  4. Borthwick A. C., Holms W. H., Nimmo H. G. The phosphorylation of Escherichia coli isocitrate dehydrogenase in intact cells. Biochem J. 1984 Sep 15;222(3):797–804. doi: 10.1042/bj2220797. [DOI] [PMC free article] [PubMed] [Google Scholar]
  5. Burnell J. N., Hatch M. D. Activation and inactivation of an enzyme catalyzed by a single, bifunctional protein: a new example and why. Arch Biochem Biophys. 1986 Mar;245(2):297–304. doi: 10.1016/0003-9861(86)90219-5. [DOI] [PubMed] [Google Scholar]
  6. Caban C. E., Ginsburg A. Glutamine synthetase adenylyltransferase from Escherichia coli: purification and physical and chemical properties. Biochemistry. 1976 Apr 6;15(7):1569–1580. doi: 10.1021/bi00652a030. [DOI] [PubMed] [Google Scholar]
  7. Casadaban M. J., Martinez-Arias A., Shapira S. K., Chou J. Beta-galactosidase gene fusions for analyzing gene expression in escherichia coli and yeast. Methods Enzymol. 1983;100:293–308. doi: 10.1016/0076-6879(83)00063-4. [DOI] [PubMed] [Google Scholar]
  8. Celenza J. L., Carlson M. A yeast gene that is essential for release from glucose repression encodes a protein kinase. Science. 1986 Sep 12;233(4769):1175–1180. doi: 10.1126/science.3526554. [DOI] [PubMed] [Google Scholar]
  9. Chung T., Klumpp D. J., LaPorte D. C. Glyoxylate bypass operon of Escherichia coli: cloning and determination of the functional map. J Bacteriol. 1988 Jan;170(1):386–392. doi: 10.1128/jb.170.1.386-392.1988. [DOI] [PMC free article] [PubMed] [Google Scholar]
  10. Cortay J. C., Bleicher F., Rieul C., Reeves H. C., Cozzone A. J. Nucleotide sequence and expression of the aceK gene coding for isocitrate dehydrogenase kinase/phosphatase in Escherichia coli. J Bacteriol. 1988 Jan;170(1):89–97. doi: 10.1128/jb.170.1.89-97.1988. [DOI] [PMC free article] [PubMed] [Google Scholar]
  11. Crawford I. P. Gene rearrangements in the evolution of the tryptophan synthetic pathway. Bacteriol Rev. 1975 Jun;39(2):87–120. doi: 10.1128/br.39.2.87-120.1975. [DOI] [PMC free article] [PubMed] [Google Scholar]
  12. Dever T. E., Glynias M. J., Merrick W. C. GTP-binding domain: three consensus sequence elements with distinct spacing. Proc Natl Acad Sci U S A. 1987 Apr;84(7):1814–1818. doi: 10.1073/pnas.84.7.1814. [DOI] [PMC free article] [PubMed] [Google Scholar]
  13. El-Maghrabi M. R., Claus T. H., Pilkis J., Fox E., Pilkis S. J. Regulation of rat liver fructose 2,6-bisphosphatase. J Biol Chem. 1982 Jul 10;257(13):7603–7607. [PubMed] [Google Scholar]
  14. Ferrara P., Duchange N., Zakin M. M., Cohen G. N. Internal homologies in the two aspartokinase-homoserine dehydrogenases of Escherichia coli K-12. Proc Natl Acad Sci U S A. 1984 May;81(10):3019–3023. doi: 10.1073/pnas.81.10.3019. [DOI] [PMC free article] [PubMed] [Google Scholar]
  15. Garcia E., Rhee S. G. Cascade control of Escherichia coli glutamine synthetase. Purification and properties of PII uridylyltransferase and uridylyl-removing enzyme. J Biol Chem. 1983 Feb 25;258(4):2246–2253. [PubMed] [Google Scholar]
  16. Garnak M., Reeves H. C. Phosphorylation of Isocitrate dehydrogenase of Escherichia coli. Science. 1979 Mar 16;203(4385):1111–1112. doi: 10.1126/science.34215. [DOI] [PubMed] [Google Scholar]
  17. Garnak M., Reeves H. C. Purification and properties of phosphorylated isocitrate dehydrogenase of Escherichia coli. J Biol Chem. 1979 Aug 25;254(16):7915–7920. [PubMed] [Google Scholar]
  18. Gilson E., Clément J. M., Brutlag D., Hofnung M. A family of dispersed repetitive extragenic palindromic DNA sequences in E. coli. EMBO J. 1984 Jun;3(6):1417–1421. doi: 10.1002/j.1460-2075.1984.tb01986.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  19. Hawke D., Yuan P. M., Shively J. E. Microsequence analysis of peptides and proteins. II. Separation of amino acid phenylthiohydantoin derivatives by high-performance liquid chromatography on octadecylsilane supports. Anal Biochem. 1982 Mar 1;120(2):302–311. doi: 10.1016/0003-2697(82)90351-7. [DOI] [PubMed] [Google Scholar]
  20. Holms W. H., Bennett P. M. Regulation of isocitrate dehydrogenase activity in Escherichia coli on adaptation to acetate. J Gen Microbiol. 1971 Jan;65(1):57–68. doi: 10.1099/00221287-65-1-57. [DOI] [PubMed] [Google Scholar]
  21. Hunter T., Cooper J. A. Protein-tyrosine kinases. Annu Rev Biochem. 1985;54:897–930. doi: 10.1146/annurev.bi.54.070185.004341. [DOI] [PubMed] [Google Scholar]
  22. KORNBERG H. L., MADSEN N. B. Synthesis of C4-dicarboxylic acids from acetate by a glyoxylate bypass of the tricarboxylic acid cycle. Biochim Biophys Acta. 1957 Jun;24(3):651–653. doi: 10.1016/0006-3002(57)90268-8. [DOI] [PubMed] [Google Scholar]
  23. Kieny M. P., Lathe R., Lecocq J. P. New versatile cloning and sequencing vectors based on bacteriophage M13. Gene. 1983 Dec;26(1):91–99. doi: 10.1016/0378-1119(83)90039-2. [DOI] [PubMed] [Google Scholar]
  24. Klionsky D. J., Skalnik D. G., Simoni R. D. Differential translation of the genes encoding the proton-translocating ATPase of Escherichia coli. J Biol Chem. 1986 Jun 25;261(18):8096–8099. [PubMed] [Google Scholar]
  25. Kornberg H. L. The role and control of the glyoxylate cycle in Escherichia coli. Biochem J. 1966 Apr;99(1):1–11. doi: 10.1042/bj0990001. [DOI] [PMC free article] [PubMed] [Google Scholar]
  26. LaPorte D. C., Chung T. A single gene codes for the kinase and phosphatase which regulate isocitrate dehydrogenase. J Biol Chem. 1985 Dec 5;260(28):15291–15297. [PubMed] [Google Scholar]
  27. LaPorte D. C., Koshland D. E., Jr A protein with kinase and phosphatase activities involved in regulation of tricarboxylic acid cycle. Nature. 1982 Dec 2;300(5891):458–460. doi: 10.1038/300458a0. [DOI] [PubMed] [Google Scholar]
  28. LaPorte D. C., Koshland D. E., Jr Phosphorylation of isocitrate dehydrogenase as a demonstration of enhanced sensitivity in covalent regulation. Nature. 1983 Sep 22;305(5932):286–290. doi: 10.1038/305286a0. [DOI] [PubMed] [Google Scholar]
  29. LaPorte D. C., Thorsness P. E., Koshland D. E., Jr Compensatory phosphorylation of isocitrate dehydrogenase. A mechanism for adaptation to the intracellular environment. J Biol Chem. 1985 Sep 5;260(19):10563–10568. [PubMed] [Google Scholar]
  30. LaPorte D. C., Walsh K., Koshland D. E., Jr The branch point effect. Ultrasensitivity and subsensitivity to metabolic control. J Biol Chem. 1984 Nov 25;259(22):14068–14075. [PubMed] [Google Scholar]
  31. Lundell D. J., Howard J. B. Isolation and sequences of the cysteinyl tryptic peptides from the MoFe-protein of Azotobacter vinelandii nitrogenase. J Biol Chem. 1981 Jun 25;256(12):6385–6391. [PubMed] [Google Scholar]
  32. Lusty C. J., Widgren E. E., Broglie K. E., Nyunoya H. Yeast carbamyl phosphate synthetase. Structure of the yeast gene and homology to Escherichia coli carbamyl phosphate synthetase. J Biol Chem. 1983 Dec 10;258(23):14466–14477. [PubMed] [Google Scholar]
  33. Maloy S. R., Nunn W. D. Genetic regulation of the glyoxylate shunt in Escherichia coli K-12. J Bacteriol. 1982 Jan;149(1):173–180. doi: 10.1128/jb.149.1.173-180.1982. [DOI] [PMC free article] [PubMed] [Google Scholar]
  34. Maxam A. M., Gilbert W. Sequencing end-labeled DNA with base-specific chemical cleavages. Methods Enzymol. 1980;65(1):499–560. doi: 10.1016/s0076-6879(80)65059-9. [DOI] [PubMed] [Google Scholar]
  35. Messing J. New M13 vectors for cloning. Methods Enzymol. 1983;101:20–78. doi: 10.1016/0076-6879(83)01005-8. [DOI] [PubMed] [Google Scholar]
  36. Neidhardt F. C., Bloch P. L., Smith D. F. Culture medium for enterobacteria. J Bacteriol. 1974 Sep;119(3):736–747. doi: 10.1128/jb.119.3.736-747.1974. [DOI] [PMC free article] [PubMed] [Google Scholar]
  37. Nimmo G. A., Nimmo H. G. The regulatory properties of isocitrate dehydrogenase kinase and isocitrate dehydrogenase phosphatase from Escherichia coli ML308 and the roles of these activities in the control of isocitrate dehydrogenase. Eur J Biochem. 1984 Jun 1;141(2):409–414. doi: 10.1111/j.1432-1033.1984.tb08206.x. [DOI] [PubMed] [Google Scholar]
  38. Ninfa A. J., Magasanik B. Covalent modification of the glnG product, NRI, by the glnL product, NRII, regulates the transcription of the glnALG operon in Escherichia coli. Proc Natl Acad Sci U S A. 1986 Aug;83(16):5909–5913. doi: 10.1073/pnas.83.16.5909. [DOI] [PMC free article] [PubMed] [Google Scholar]
  39. Norrander J., Kempe T., Messing J. Construction of improved M13 vectors using oligodeoxynucleotide-directed mutagenesis. Gene. 1983 Dec;26(1):101–106. doi: 10.1016/0378-1119(83)90040-9. [DOI] [PubMed] [Google Scholar]
  40. Poorman R. A., Randolph A., Kemp R. G., Heinrikson R. L. Evolution of phosphofructokinase--gene duplication and creation of new effector sites. 1984 May 31-Jun 6Nature. 309(5967):467–469. doi: 10.1038/309467a0. [DOI] [PubMed] [Google Scholar]
  41. Reed S. I., Hadwiger J. A., Lörincz A. T. Protein kinase activity associated with the product of the yeast cell division cycle gene CDC28. Proc Natl Acad Sci U S A. 1985 Jun;82(12):4055–4059. doi: 10.1073/pnas.82.12.4055. [DOI] [PMC free article] [PubMed] [Google Scholar]
  42. Reimann E. M., Titani K., Ericsson L. H., Wade R. D., Fischer E. H., Walsh K. A. Homology of the gamma subunit of phosphorylase b kinase with cAMP-dependent protein kinase. Biochemistry. 1984 Aug 28;23(18):4185–4192. doi: 10.1021/bi00313a027. [DOI] [PubMed] [Google Scholar]
  43. Sanger F., Coulson A. R. A rapid method for determining sequences in DNA by primed synthesis with DNA polymerase. J Mol Biol. 1975 May 25;94(3):441–448. doi: 10.1016/0022-2836(75)90213-2. [DOI] [PubMed] [Google Scholar]
  44. Shoji S., Ericsson L. H., Walsh K. A., Fischer E. H., Titani K. Amino acid sequence of the catalytic subunit of bovine type II adenosine cyclic 3',5'-phosphate dependent protein kinase. Biochemistry. 1983 Jul 19;22(15):3702–3709. doi: 10.1021/bi00284a025. [DOI] [PubMed] [Google Scholar]
  45. Stern M. J., Ames G. F., Smith N. H., Robinson E. C., Higgins C. F. Repetitive extragenic palindromic sequences: a major component of the bacterial genome. Cell. 1984 Jul;37(3):1015–1026. doi: 10.1016/0092-8674(84)90436-7. [DOI] [PubMed] [Google Scholar]
  46. Takio K., Wade R. D., Smith S. B., Krebs E. G., Walsh K. A., Titani K. Guanosine cyclic 3',5'-phosphate dependent protein kinase, a chimeric protein homologous with two separate protein families. Biochemistry. 1984 Aug 28;23(18):4207–4218. doi: 10.1021/bi00313a030. [DOI] [PubMed] [Google Scholar]
  47. Thorsness P. E., Koshland D. E., Jr Inactivation of isocitrate dehydrogenase by phosphorylation is mediated by the negative charge of the phosphate. J Biol Chem. 1987 Aug 5;262(22):10422–10425. [PubMed] [Google Scholar]
  48. Tinoco I., Jr, Borer P. N., Dengler B., Levin M. D., Uhlenbeck O. C., Crothers D. M., Bralla J. Improved estimation of secondary structure in ribonucleic acids. Nat New Biol. 1973 Nov 14;246(150):40–41. doi: 10.1038/newbio246040a0. [DOI] [PubMed] [Google Scholar]
  49. Tinoco I., Jr, Uhlenbeck O. C., Levine M. D. Estimation of secondary structure in ribonucleic acids. Nature. 1971 Apr 9;230(5293):362–367. doi: 10.1038/230362a0. [DOI] [PubMed] [Google Scholar]
  50. Wang J. Y., Koshland D. E., Jr The reversible phosphorylation of isocitrate dehydrogenase of Salmonella typhimurium. Arch Biochem Biophys. 1982 Oct 1;218(1):59–67. doi: 10.1016/0003-9861(82)90321-6. [DOI] [PubMed] [Google Scholar]
  51. Yanisch-Perron C., Vieira J., Messing J. Improved M13 phage cloning vectors and host strains: nucleotide sequences of the M13mp18 and pUC19 vectors. Gene. 1985;33(1):103–119. doi: 10.1016/0378-1119(85)90120-9. [DOI] [PubMed] [Google Scholar]

Articles from Journal of Bacteriology are provided here courtesy of American Society for Microbiology (ASM)

RESOURCES