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. 1992 Sep;174(17):5748–5752. doi: 10.1128/jb.174.17.5748-5752.1992

Cloning and sequencing of Escherichia coli murZ and purification of its product, a UDP-N-acetylglucosamine enolpyruvyl transferase.

J L Marquardt 1, D A Siegele 1, R Kolter 1, C T Walsh 1
PMCID: PMC206525  PMID: 1512209

Abstract

The Escherichia coli gene murZ, encoding the enzyme UDP-N-acetylglucosamine enolpyruvyl transferase, has been cloned and sequenced. Identified by screening an E. coli genomic library for clones that conferred phosphomycin resistance, murZ encoded a 419-amino-acid polypeptide and was mapped to 69.3 min on the E. coli chromosome. MurZ protein was purified to near homogeneity and found to have the expected UDP-N-acetylglucosamine enolpyruvyl transferase activity. Sequence analysis of the predicted product revealed 44% identity to OrfR from Bacillus subtilis (K. Trach, J.W. Chapman, P. Piggot, D. LeCoq, and J.A. Hoch, J. Bacteriol. 170:4194-4208, 1988), suggesting that orfR may also encode a UDP-N-acetylglucosamine enolpyruvyl transferase enzyme. MurZ is also homologous to the aromatic amino acid biosynthetic enzyme enolpyruvyl shikimate phosphate synthase, the other enzyme known to catalyze an enolpyruvyl transfer.

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Selected References

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  1. Alper M. D., Ames B. N. Transport of antibiotics and metabolite analogs by systems under cyclic AMP control: positive selection of Salmonella typhimurium cya and crp mutants. J Bacteriol. 1978 Jan;133(1):149–157. doi: 10.1128/jb.133.1.149-157.1978. [DOI] [PMC free article] [PubMed] [Google Scholar]
  2. Altschul S. F., Gish W., Miller W., Myers E. W., Lipman D. J. Basic local alignment search tool. J Mol Biol. 1990 Oct 5;215(3):403–410. doi: 10.1016/S0022-2836(05)80360-2. [DOI] [PubMed] [Google Scholar]
  3. Cassidy P. J., Kahan F. M. A stable enzyme-phosphoenolpyruvate intermediate in the synthesis of uridine-5'-diphospho-N-acetyl-2-amino-2-deoxyglucose 3-O-enolpyruvyl ether. Biochemistry. 1973 Mar 27;12(7):1364–1374. doi: 10.1021/bi00731a017. [DOI] [PubMed] [Google Scholar]
  4. Choi Y. L., Nishida T., Kawamukai M., Utsumi R., Sakai H., Komano T. Cloning and sequencing of an Escherichia coli gene, nlp, highly homologous to the ner genes of bacteriophages Mu and D108. J Bacteriol. 1989 Sep;171(9):5222–5225. doi: 10.1128/jb.171.9.5222-5225.1989. [DOI] [PMC free article] [PubMed] [Google Scholar]
  5. Guyer M. S. The gamma delta sequence of F is an insertion sequence. J Mol Biol. 1978 Dec 15;126(3):347–365. doi: 10.1016/0022-2836(78)90045-1. [DOI] [PubMed] [Google Scholar]
  6. Guyer M. S. Uses of the transposon gamma delta in the analysis of cloned genes. Methods Enzymol. 1983;101:362–369. doi: 10.1016/0076-6879(83)01027-7. [DOI] [PubMed] [Google Scholar]
  7. Kahan F. M., Kahan J. S., Cassidy P. J., Kropp H. The mechanism of action of fosfomycin (phosphonomycin). Ann N Y Acad Sci. 1974 May 10;235(0):364–386. doi: 10.1111/j.1749-6632.1974.tb43277.x. [DOI] [PubMed] [Google Scholar]
  8. Lanzetta P. A., Alvarez L. J., Reinach P. S., Candia O. A. An improved assay for nanomole amounts of inorganic phosphate. Anal Biochem. 1979 Nov 15;100(1):95–97. doi: 10.1016/0003-2697(79)90115-5. [DOI] [PubMed] [Google Scholar]
  9. Liu L., Whalen W., Das A., Berg C. M. Rapid sequencing of cloned DNA using a transposon for bidirectional priming: sequence of the Escherichia coli K-12 avtA gene. Nucleic Acids Res. 1987 Nov 25;15(22):9461–9469. doi: 10.1093/nar/15.22.9461. [DOI] [PMC free article] [PubMed] [Google Scholar]
  10. MacFerrin K. D., Terranova M. P., Schreiber S. L., Verdine G. L. Overproduction and dissection of proteins by the expression-cassette polymerase chain reaction. Proc Natl Acad Sci U S A. 1990 Mar;87(5):1937–1941. doi: 10.1073/pnas.87.5.1937. [DOI] [PMC free article] [PubMed] [Google Scholar]
  11. Maskell D. J., Morrissey P., Dougan G. Cloning and nucleotide sequence of the aroA gene of Bordetella pertussis. J Bacteriol. 1988 Jun;170(6):2467–2471. doi: 10.1128/jb.170.6.2467-2471.1988. [DOI] [PMC free article] [PubMed] [Google Scholar]
  12. Stallings W. C., Abdel-Meguid S. S., Lim L. W., Shieh H. S., Dayringer H. E., Leimgruber N. K., Stegeman R. A., Anderson K. S., Sikorski J. A., Padgette S. R. Structure and topological symmetry of the glyphosate target 5-enolpyruvylshikimate-3-phosphate synthase: a distinctive protein fold. Proc Natl Acad Sci U S A. 1991 Jun 1;88(11):5046–5050. doi: 10.1073/pnas.88.11.5046. [DOI] [PMC free article] [PubMed] [Google Scholar]
  13. Trach K., Chapman J. W., Piggot P., LeCoq D., Hoch J. A. Complete sequence and transcriptional analysis of the spo0F region of the Bacillus subtilis chromosome. J Bacteriol. 1988 Sep;170(9):4194–4208. doi: 10.1128/jb.170.9.4194-4208.1988. [DOI] [PMC free article] [PubMed] [Google Scholar]
  14. Venkateswaran P. S., Wu H. C. Isolation and characterization of a phosphonomycin-resistant mutant of Escherichia coli K-12. J Bacteriol. 1972 Jun;110(3):935–944. doi: 10.1128/jb.110.3.935-944.1972. [DOI] [PMC free article] [PubMed] [Google Scholar]
  15. Wanke C., Falchetto R., Amrhein N. The UDP-N-acetylglucosamine 1-carboxyvinyl-transferase of Enterobacter cloacae. Molecular cloning, sequencing of the gene and overexpression of the enzyme. FEBS Lett. 1992 Apr 27;301(3):271–276. doi: 10.1016/0014-5793(92)80255-f. [DOI] [PubMed] [Google Scholar]
  16. Waterman M. S., Eggert M. A new algorithm for best subsequence alignments with application to tRNA-rRNA comparisons. J Mol Biol. 1987 Oct 20;197(4):723–728. doi: 10.1016/0022-2836(87)90478-5. [DOI] [PubMed] [Google Scholar]
  17. Wu H. C., Venkateswaran P. S. Fosfomycin-resistant mutant of Escherichia coli. Ann N Y Acad Sci. 1974 May 10;235(0):587–592. doi: 10.1111/j.1749-6632.1974.tb43292.x. [DOI] [PubMed] [Google Scholar]
  18. 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]

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