1887

Abstract

DNA fragments from and were isolated which restored hydrogenase activities in both and mutant strains of The and genes, which map near minute 59 of the genome map, 17 kb distant from each other, are not structural hydrogenase genes, but mutation in either of these genes leads to failure to synthesize any of the hydrogenase isoenzymes. The smallest DNA fragments which restored hydrogenase activity to both mutant strains were 4·7 kb from and 2·3 kb from These fragments were cleaved into smaller fragments which did not complement either of the mutations. The cloned heterologous genes also restored formate hydrogenlyase activity but they did not restore activity in or mutant strains of The cloned genes, on plasmids, did not lead to the synthesis of proteins of sufficient size to be the hydrogenase catalytic subunit. The hydrogenase proteins synthesized by and mutant strains of transformed by cloned genes from and were shown by isoelectric and immunological methods to be hydrogenase. Thus, these genes are not hydrogenase structural genes.

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1990-06-01
2024-04-25
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References

  1. Ballantine S.P., Boxer D.H. 1985; Nickel-containing hydrogenaseisoenzymes from anaerobically grown Escherichia coli K12. Journal of Bacteriology 163:454–459
    [Google Scholar]
  2. Ballantine S.P., Boxer D.H. 1986; Isolation and characterization of a soluble active fragment of hydrogenaseisoenzyme 2 from the membranes of anaerobically grown Escherichia coli. European Journal of Biochemistry 156:277–284
    [Google Scholar]
  3. Blake M.S., Johnston K.H., Russell-Jones G.J., Gotschlich E.C. 1984; A rapid, sensitive method for detection of alkaline phosphatase-conjugated anti-antibody on western blots. Analytical Biochemistry 136:175–179
    [Google Scholar]
  4. Chaudhuri A., Krasna A.I. 1987; Isolation of genes required for hydrogenase synthesis in Escherichia coli. Journal of General Microbiology 133:3289–3298
    [Google Scholar]
  5. Chaudhuri A., Krasna A.I. 1988; Isolation of a gene required for growth of Escherichia coli on fumarate and H2. Journal of General Microbiology 134:2155–2164
    [Google Scholar]
  6. Gitlitz P., Krasna A.I. 1975; Structural and catalytic properties of hydrogenase from Chromatium. Biochemistry 14:2561–2568
    [Google Scholar]
  7. Harker A.R., Zuber M., Evans H.J. 1986; Immunological homology between the membrane-bound hydrogenase of Rhizobium japonicum and Escherichia coli. Journal of Bacteriology 165:579–584
    [Google Scholar]
  8. Kanayama H., Urano N., Aihara C., Karube I. 1986; Cloning and expression of Citrobacterfreundiihydrogenase genes in Escherichia coli. Applied Microbiology and Biotechnology 24:392–396
    [Google Scholar]
  9. Karube I., Urano N., Yamada T., Hirochika H., Sakaguchi K. 1983; Cloning and expression of the hydrogenase gene from Clostridium butyricum in Escherichia coli. FEBS tetters 158:119–122
    [Google Scholar]
  10. Karube I., Tomiyama A., Kikuchi A. 1984; Molecular cloning and physical mapping of the hyd gene of Escherichia coli K-12. FEMS Microbiology tetters 25:165–168
    [Google Scholar]
  11. Kohara Y., Kiyotaka A., Katsumi I. 1987; The physical map of the whole E. coli chromosome; application of a new strategy for rapid analysis and sorting of a large genomic library. Cell 50:495–508
    [Google Scholar]
  12. Lee J.H., Patel P., Sankar P., Shanmugam K.T. 1985; Isolation and characterization of mutant strains of Escherichia coli altered in H2 metabolism. Journal of Bacteriology 162:344–352
    [Google Scholar]
  13. Sancar A., Hack A.M., Rupp W.D. 1979; Simple method for identification of plasmid-coded proteins. Journal of Bacteriology 137:692–693
    [Google Scholar]
  14. Sankar P., Lee J.H., Shanmugam K.T. 1985; Cloning of hydrogenase genes and fine structure analysis of an operon essential for H2 metabolism in Escherichia coli. Journal of Bacteriology 162:353–360
    [Google Scholar]
  15. Sankar P., Shanmugam K.T. 1988a; Biochemical and genetic analysis of hydrogen metabolism in Escherichia coli: the hydB gene. Journal of Bacteriology 170:5433–5439
    [Google Scholar]
  16. Sankar P., Shanmugam K.T. 1988b; Hydrogen metabolism in Escherichia coli: biochemical and genetic evidence for a hydF gene. Journal of Bacteriology 170:5446–5451
    [Google Scholar]
  17. Sankar P., Lee J.H., Shanmugam K.T. 1988; Gene-product relationships of fhl and fdv genes of Escherichia coli. Journal of Bacteriology 170:5440–5445
    [Google Scholar]
  18. Sawers R.G., Ballantine S.P., Boxer D.H. 1985; Differential expression of hydrogenaseisoenzymes in Escherichia coli K12: evidence for a third isoenzyme. Journal of Bacteriology 164:1324–1331
    [Google Scholar]
  19. Sawers R.G., Boxer D.H. 1986; Purification and properties of membrane-bound hydrogenaseisoenzyme 1 from anaerobically grown Escherichia coli K12. European Journal of Biochemistry 156:265–275
    [Google Scholar]
  20. Schengrund C., Krasna A.I. 1969; Purification and properties of the light-activated hydrogenase of Proteus vulgaris. Biochimica et Biophysica Acta 185:332–337
    [Google Scholar]
  21. Waugh R., Boxer D.H. 1986; Pleiotropic hydrogenase mutants of Escherichia coli K12: growth in the presence of nickel can restore hydrogenase activity. Biochimie 68:157–166
    [Google Scholar]
  22. Wu L.F., Mandrand-Berthelot M.-A. 1986; Genetic and physiological characterization of new Escherichia coli mutants impaired in hydrogenase activity. Biochimie 68:167–179
    [Google Scholar]
  23. Yerkes J.H., Casson L.P., Honkanen A.K., Walker G.C. 1984; Anaerobiosis induces expression of ant, a new Escherichia coli locus with a role in anaerobic electron transport. Journal of Bacteriology 158:180–186
    [Google Scholar]
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