Molecular and General Genetics MGG

, Volume 247, Issue 5, pp 546–554 | Cite as

Characterization of three genes in the dam-containing operon of Escherichia coli

  • Anita Lyngstadaas
  • Anders Løbner-Olesen
  • Erik Boye
Original Paper

Abstract

The dam-containing operon in Escherichia coli is located at 74 min on the chromosomal map and contains the genes aroK, aroB, a gene called urf74.3, dam and trpS. We have determined the nucleotide sequence between the dam and trpS genes and show that it encodes two proteins with molecular weights of 24 and 27 kDa. Furthermore, we characterize the three genes urf74.3, 24kDa, 27kDa and the proteins they encode. The predicted amino acid sequences of the 24 and 27 kDa proteins are similar to those of the CbbE and CbbZ proteins, respectively, of the Alcaligenes eutrophus cbb operon, which encodes enzymes involved in the Calvin cycle. In separate experiments, we have shown that the 24 kDa protein has d-ribulose-5-phosphate epimerase activity (similar to CbbE), and we call the gene rpe. Similarly, the 27 kDa protein has 2-phosphoglycolate phosphatase activity (similar to CbbZ), and we name the gene gph. The Urf74.3 protein, with a predicted molecular weight of 46 kDa, migrated as a 70 kDa product under denaturing conditions. Overexpression of Urf74.3 induced cell filamentation, indicating that Urf74.3 directly or indirectly interferes with cell division. We present evidence for translational coupling between aroB and urf74.3 and also between rpe and gph. Proteins encoded in the dam superoperon appear to be largely unrelated: Dam, and perhaps Urf74.3, are involved in cell cycle regulation, AroK, AroB, and TrpS function in aromatic amino acid biosynthesis, whereas Rpe and Gph are involved in carbohydrate metabolism.

Key words

dam-containing operon Translational coupling Escherichia coli Ribulose-5-phosphate epimerase 2-Phosphoglycolate phosphatase 

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References

  1. Bachmann BJ (1990) Linkage map of Escherichia coli K-12, Edition 8. Microbiol Rev 54:130–197Google Scholar
  2. Barras F, Marinus MG (1989) The great GATC: DNA methylation in E. coli. Trends Genet 5: 139–143Google Scholar
  3. Berlin B (1993) Studies on the activation of E. coli σ70 promoters by CRP and AraC. Ph.D. thesis, University of HeidelbergGoogle Scholar
  4. Boye E, Løbner-Olesen A (1990) The role of dam methyltransferase in the control of DNA replication in Escherichia coli. Cell 62:981–989Google Scholar
  5. Boye E, Løbner-Olesen A (1991) Bacterial growth studied by flow cytometry. Res Microbiol 142:131–135Google Scholar
  6. Brooks JE, Blumenthal RM, Gingeras TR (1983) The isolation and characterization of the Escherichia coli DNA adenine methylase (dam) gene. Nucleic Acids Res 11: 837–851Google Scholar
  7. Campbell JL, Kleckner N (1990) E. coli oriC and dnaA gene promoter are sequestered from dam methyltransferase following the passage of the chromosomal replication fork. Cell 62: 967–979Google Scholar
  8. Casabadan MJ, Cohen SN (1980) Analysis of gene control signals by DNA fusion and cloning in Escherichia coli. J Mol Biol 138:99–112Google Scholar
  9. Casaregola S, Jacq A, Laoudj D, McGurk G, Margarson S, Tempete M, Norris V, Holland IB (1992) Cloning and analysis of the entire Escherichia coli ams gene: ams is identical to hmpl and encodes a 114 kDa protein that migrates as a 180 kDa protein. J Mol Biol 228:30–40Google Scholar
  10. Clark DJ, Maaløe O (1967) DNA replication and the division cycle in Escherichia coli. J Mol Biol 23:99–112Google Scholar
  11. Dar ME, Bhagwat AS (1993) Mechanism of expression of DNA repair gene vsr, an Escherichia coli gene that overlaps the DNA cytosine methylase gene, dcm. Mol Microbiol 9: 823–833Google Scholar
  12. Essar DW, Eberly L, Crawford IP (1990) Evolutionary differences in chromosomal locations of four early genes of the trypophan pathway in fluorescent Pseudomonas: DNA sequences and characterization of Pseudomonas putida trpE and trpGDC. J Bacteriol 172:867–883Google Scholar
  13. Hall CV, Yanofsky C (1981) The nucleotide sequence of the structural gene for Escherichia coli tryptophanyl-tRNA synthetase. J Bacteriol 148:941–949Google Scholar
  14. Hall CV, vanCleemputt M, Muench KH, Yanofsky C (1982) The nucleotide sequence of the structural gene for Escherichia coli tryptophanyl-tRNA synthetase. J Biol Chem 257: 6132–6136Google Scholar
  15. Hattman SM, Brooks JE, Masurekar M (1978) Sequence specificity of the PI modification methylase (M.Eco P1) and the DNA methylase (M.Eco dam) contolled by the E. coli dam gene. J Mol Biol 126:367–380Google Scholar
  16. Herman GE, Modrich P (1981) Escherichia coli K-12 clones that overproduce dam methylase are hypermutable. J Bacteriol 145:644–646Google Scholar
  17. Jonczyk P, Hines R, Smith DW (1989) The Escherichia coli dam gene is expressed as a distal gene of a new operon. Mol Gen Genet 217:85–96Google Scholar
  18. Kushner SR, Nagaishi H, Templin A, Clark AJ (1971) Genetic recombination in Escherichia coli: the role of exonuclease I. Proc Natl Acad Sci USA 68:824–825Google Scholar
  19. Lacks S, Greenberg B (1977) Complementary specificity of restriction endonucleases of Diplococcus pneumoniae with respect to DNA methylation. J Mol Biol 114:153–168Google Scholar
  20. Laemmli UK (1970) Cleavage of structural proteins during the assembly of the head of bacteriophage T4. Nature 227: 680–685Google Scholar
  21. Larsen JEL, Gerdes K, Light J, Molin S (1984) Low-copy-number plasmid-cloning vectors amplifiable by derepression of an inserted foreign promoter. Gene 28:45–54Google Scholar
  22. Løbner-Olesen A, Marinus MG (1992) Identification of the gene (aroK) encoding shikimic acid kinase I of Escherichia coli. J Bacteriol 174: 525–529Google Scholar
  23. Løbner-Olesen A, Boye E, Marinus MG (1992) Expression of the Escherichia coli dam gene. Mol Microbiol 6:1841–1851Google Scholar
  24. Macdonald H, Cole J (1985) Molecular cloning and functional analysis of the cysG and nirB genes of Escherichia coli K-12, two closely-linked genes required for NADPH-dependent nitrite reductase activity. Mol Gen Genet 200:328–334Google Scholar
  25. Marinus MG (1973) Location of DNA methylation genes on the Escherichia coli genetic map. Mol Gen Genet 127:47–55Google Scholar
  26. Millar G, Coggins JR (1986) The complete amino acid sequence of 3-dehydroquinate synthase of Escherichia coli K-12. FEBS Lett 200: 11–17Google Scholar
  27. Miller JF (1992). A short course in bacterial genetics. Cold Spring Harbor Laboratory Press, Cold Spring Harbor, New York, USAGoogle Scholar
  28. Oka A, Sugisaki H, Taganami M (1981) Nucleotide sequence of the kanamycin resistance transposon Tn903. J Mol Biol 147:217–226Google Scholar
  29. Pearson WR, Lipman DJ (1988) Improved tools for biological sequence comparison. Proc Natl Acad Sci USA 85:2444–2448Google Scholar
  30. Rasmussen LJ, Møller PL, Atlung T (1991) Carbon metabolism regulates expression of the pfl (pyruvate formate-lyase) gene in Escherichia coli. J Bacteriol 173: 6390–6397Google Scholar
  31. Rasmussen LJ, Marinus MG, Løbner-Olesen A (1994) Novel growth rate control of dam gene expression in E. coli. Mol Microbiol 12:631–638Google Scholar
  32. Russel DW, Zinder ND (1987) Hemimethylation prevents DNA replication in E. coli. Cell 50: 1071–1079Google Scholar
  33. Sancar A, Hack AM, Rupp WD (1979) Simple method for identification of plasmid-coded proteins. J Bacteriol 137:692–693Google Scholar
  34. Sanger F, Nicklen S, Coulson AR (1977) DNA sequencing with chain terminating inhibitors. Proc Natl Acad Sci USA 74:5463–5467Google Scholar
  35. Schäferjohann J, Yoo J-G, Kusian, B, Bowien B (1993) The cbb operons of the facultative chemoautotroph Alcaligenes eutrophus encode phosphoglycolate phosphatase. J Bacteriol 175:7329–7340Google Scholar
  36. Schümperli D, McKenney K, Sobieski DA, Rosenberg M (1982) Translational coupling at an intercistronic boundary of the Escherichia coli galactose operon. Cell 30: 865–871Google Scholar
  37. Skarstad K, Steen HB, Boye E (1985) Timing and initiation of chromosome replication in individual Escherichia coli cells. J Bacteriol 163:661–668Google Scholar
  38. Southern E (1975) Detection of specific sequences among DNA fragments separated by gel electrophoresis. J Mol Biol 98:503–517Google Scholar
  39. Tsui H-CT, Zhao G, Feng G, Leung H-CE, Winkler ME (1994) The mutL repair gene of Escherichia coli K-12 forms a superoperon with a gene encoding a new cell-wall amidase. Mol Microbiol 11: 189–202Google Scholar
  40. Wu T-H, Grelland E, Boye E, Marinus MG (1992) Identification of a weak promoter for the dam gene of Escherichia coli. Biochim Biophys Acta 1131:47–52Google Scholar
  41. Yanisch-Perron C, Vieira J, Messing J (1985) Improved M13 phage containing vectors and host strains: nucleotide sequences of the M13mpl8 and pUC19 vectors. Gene 33:103–119Google Scholar
  42. Zubay G (1980) The isolation and properties of CAP, the catabolite gene activator. Methods Enzymol 65: 856–877Google Scholar

Copyright information

© Springer-Verlag 1995

Authors and Affiliations

  • Anita Lyngstadaas
    • 1
  • Anders Løbner-Olesen
    • 2
  • Erik Boye
    • 1
  1. 1.Department of BiophysicsInstitute for Cancer ResearchOsloNorway
  2. 2.Department of Molecular Cell BiologyUniversity of CopenhagenCopenhagenDenmark

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