Nucleotide sequence and expression analysis of the phosphoglucomutase gene Free

Abstract

The gene () encoding phosphoglucomutase (EC 5·4.2·2) has previously been cloned by complementation of cellulose-negative mutants. In the present report the nucleotide sequence of a 2·0 kb DNA fragment containing is described. Expression analysis using the bacteriophage T7 RNA polymerase promoter 10 resulted in identification of a probable translational start codon of , and this conclusion was confirmed by N-terminal amino acid sequencing of the recombinant protein. From the nucleotide sequence data it was deduced that encodes a protein with a calculated molecular mass of 59·6 kDa. A protein of similar size was visualized after transcription and translation, using the cloned 2·0 kb fragment as template. The results of an amino acid sequence comparison and a biochemical analysis indicated that the CelB protein is structurally and functionally related to the previously characterized human and rabbit phosphoglucomutases.

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1994-05-01
2024-03-28
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References

  1. Adhya S., Schwartz M. 1971; Phosphoglucose mutants of Escherichia coli K-12. J Bacteriol 108:621–626
    [Google Scholar]
  2. Altschul S. F., Gish W., Miller W., Myers E. W., Lipman D. J. 1990; Basic local alignment search tool. J Mol Biol 215:403–410
    [Google Scholar]
  3. Chung C. T., Niemela S. L., Miller R. H. 1989; One-step preparation of competent Escherichia coli: transformation and storage of bacterial cells in the same solution. Proc Natl Acad Sci USA 86:2172–2175
    [Google Scholar]
  4. Dai J.-B., Liu Y., Ray W. J., Konno M. 1992; The crystal structure of muscle phosphoglucomutase refined at 2,7-angstrom resolution. J Biol Chem 267:6322–6337
    [Google Scholar]
  5. Fjaervik E., Frydenlund K., Valla S., Huggirat Y., Benziman M. 1991; Complementation of cellulose-negative mutants of Aceto- bacter xylinum by the cloned structural gene for phosphoglucomutase. FEMS Microbiol Lett 77:325–330
    [Google Scholar]
  6. Henikoff S. 1987; Unidirectional digestion with exonuclease III in DNA sequence analysis. Methods Enyymol 155:156–165
    [Google Scholar]
  7. Jiang X. M., Neal B., Santiago F., Lee S. J., Romana L. K., Reeves P. R. 1991; Structure and sequence of the rfb (O antigen) gene cluster of Salmonella serovar typhimurium (strain LT2). Mol Microbiol 5:695–713
    [Google Scholar]
  8. Keen N. T., Tamaki S., Kobayashi D., Trollinger D. 1988; Improved broad-host-range plasmids for DNA cloning in Gram-negative bacteria. Gene 70:191–197
    [Google Scholar]
  9. Koplin R., Arnold B., Hotte B., Simon R., Wang G., Piihler A. 1992; Genetics of xanthan production in Xanthomonas campestris: the xan A and xan B genes are involved in UDP-glucose and GDP-mannose biosynthesis. J Bacteriol 174:191–199
    [Google Scholar]
  10. Lowry O.H., Passonneau J. W. 1969; Phosphoglucomutase kinetics with the phosphates of fructose, glucose, mannose, ribose, and galactose. J Biol Chem 244:910–916
    [Google Scholar]
  11. Marolda C.L., Valvano M. A. 1993; Identification, expression, and DNA sequence of the GDP-mannose biosynthesis genes encoded by the 07 rfb gene cluster of strain vW 187 (Escherichia coli 07 :K1). J Bacteriol 175:148–158
    [Google Scholar]
  12. Ray W. J., Peck E. J. 1972; Phosphomutases.. In The Enzymes, 3rd edn. 6 pp. 407–458 Edited by Boyer P. D. New York: Academic Press;
    [Google Scholar]
  13. Ray W. J., Hermodson M. A., Puvathingal J. M., Mahoney W. C. 1983; The complete amino acid sequence of rabbit muscle phosphoglucomutase. J Biol Chem 258:9166–9174
    [Google Scholar]
  14. Rhyu G. I., Ray W. J., Markley J. L. 1984; Enzyme-bound intermediates in the conversion of glucose-1-phosphate to glucose- 6-phosphate by phosphoglucomutase. Phosphorus NMR studies. Biochemistry 23:252–260
    [Google Scholar]
  15. Sa'-Correia I., Darzins A., Wang S. K., Berry A., Chakrabarty A. M. 1987; Alginate biosynthetic enzymes in mucoid and nonmucoid Pseudomonas aeruginosa: overproduction of phospho- mannose isomerase, phosphomannomutase, and GDP-mannose pyrophosphorylase by overexpression of the phosphomannose isomerase [pmi) gene. J Bacteriol 169:3224–3231
    [Google Scholar]
  16. Sambrook J., Fritsch E. F., Maniatis T. 1989 Molecular Cloning: a Eaboratory Manual, 2 nd edn. Cold Spring Harbor, NY: Cold Spring Harbor Laboratory;
    [Google Scholar]
  17. Sanger F., Nicklen S., Coulson A. R. 1977; DNA sequencing with chain-terminating inhibitors. Proc Natl Acad Sci USA 74:5463–5467
    [Google Scholar]
  18. Stevenson G., Lee S. J., Romana L. K., Reeves P. R. 1991; The cps gene cluster of Salmonella strain LT2 includes a second mannose pathway: sequence of two genes and relationship to genes in the rfb gene cluster. Mol and Gen Genet 227:173–180
    [Google Scholar]
  19. Tabor S., Richardson C. C. 1985; A bacteriophage T7 RNA polymerase/promoter system for controlled exclusive expression of specific genes. Proc Natl Acad Sci USA 82:1074–1078
    [Google Scholar]
  20. Valla S., Coucheron D. H., Fjaervik E., Kjosbakken J., Weinhouse H., Ross P., Amikan D., Benziman M. 1989; Cloning of a gene involved in cellulose biosynthesis in Acetobacter xylinum: complementation of cellulose-negative mutants by the UDPG pyrophosphorylase structural gene.. Mol J Gen Genet 217:26–30
    [Google Scholar]
  21. Whitehouse D. B., Putt W., Lovegrove J. U., Morrison K., Hollyoake M., Fox M. F., Hopkinson D. A., Edwards Y. H. 1992; Phosphoglucomutase 1: complete human and rabbit mRNA sequences and direct mapping of this highly polymorphic marker on human chromosome 1. Proc Natl Acad Sci USA 89:411–415
    [Google Scholar]
  22. Zielinski N. A., Chakrabarty A. M., Berry A. 1991; Characterization and regulation of the Pseudomonas aeruginosa alg C gene encoding phosphomannomutase. J Biol Chem 266:9754–9763
    [Google Scholar]
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