1887

Abstract

Mutants deficient in the proper regulation and derepression of ribulose-1,5-bisphosphate carboxylase/oxygenase (RuBPC/O) in were isolated by ethyl methanesulphonate (EMS) and Tn5 mutagenesis of a parental strain. Mutants were identified by their ability to grow under conditions where the organism requires basal levels of RuBPC/O for growth yet fail to grow under conditions which require derepression of the enzyme (Aut). The newly isolated Aut mutants exhibited phenotypes distinguishable from the previously isolated Aut mutant, strain KW25/11. Rocket immunoelectrophoretic examination of RuBPC/O levels revealed marked variance in the ability of mutants to derepress form I and form II RuBPC/O in the absence of exogenous carbon. Evidence that some of the mutants possessed different mutations was substantiated by complementation of the EMS-generated mutants by entirely different genes isolated from a genomic library of constructed in the broad-host-range cosmid vector pVK102. Southern hybridization analysis of the complementing library isolates showed the complementing genes to be normally carried on the endogenous plasmids of . The gene complementing mutant strain KW25/11 was mapped by Tn5 insertional inactivation and the complementing region found to reside on a 1·5 kb I-HI fragment. Complemented strains were unable to match wild-type levels of RuBPC/O under conditions requiring derepression of the enzyme, except for mutant strain EMS45. The Aut phenotype, represented by the mutants isolated in this study, stems from a deficiency in some aspect of photoautotrophic growth.

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1989-06-01
2021-10-23
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References

  1. Andersen K., Valentine R.C. 1982 Genetic Engineering of Microorganisms for Chemicals, pp 446–458 Hollaender A., De Moss R. D., Kaplan S., Konisky J., Savage D., Wolfe R. S. Edited by New York: Plenum Press;
    [Google Scholar]
  2. Andersen K., Wilke-Douglas M. 1984; Construction and use of a gene bank of Alcalignes eutrophus in the analysis of ribulose bisphosphate carboxylase genes. Journal of Bacteriology 159:973–978
    [Google Scholar]
  3. Andersen K., Wilke-Douglas M. 1987; Genetic and physical mapping and expression in Pseudomonas aeruginosa of the chromosomally encoded ribulose bisphosphate carboxylase gene of Alcali- genes eutropus.. Journal of Bacteriology 169:1997–2004
    [Google Scholar]
  4. Berg D.E., Schmandt M.A., Lowe J.B. 1983; Specificity of transposon Tn5 insertion. Genetics 105:813–828
    [Google Scholar]
  5. Birnboim H.C., Doly J. 1979; A rapid alkaline extraction procedure for screening recombinant plasmid DNA. Nucleic Acids Research 7:1513–1517
    [Google Scholar]
  6. Bowien B., Friedrich B., Friedrich C.G. 1984; Involvement of megaplasmids in heterotrophic derepression of the carbon dioxide assimilating enzyme system in Alcaligenes spp. Archives of Microbiology 139:305–310
    [Google Scholar]
  7. Boyer H.B., Roulland-Sussaiz D. 1969; A complementation analysis of the restriction and modification of DNA in Escherichia coli.. Journal of Molecular Biology 41:459–472
    [Google Scholar]
  8. Cohen S.N., Chang A.C.Y., Hsu L. 1972; Nonchromosomal antibiotic resistance in bacteria: genetic transformation of Escherichia coli by R factor DNA. Proceedings of the National Academy of Sciences of the United States of America 69:2110–2114
    [Google Scholar]
  9. Ditta G., Schmidhauser T., Yakoboson E., Lu P., Liang X., Finaly D., Guiney D., Helinski D.R. 1985; Plasmids related to the broad host range vector pRK290 useful for gene cloning and for monitoring gene expression. Plasmid 13:149–153
    [Google Scholar]
  10. Dow C.S. 1987; CO2 fixation in Rhodopseudomonas blastica.. In Microbial Growth on C1 Compounds, pp 28–37 Van Verseld H. W., Duine J. A. Edited by Dordrecht: Martinus Nijhoff;
    [Google Scholar]
  11. Falcone D.L., Quivey R.G.Jr Tabita F.R. 1988; Transposon mutagenesis and physiological analysis of strains containing inactivated form I and form II ribulose bisphosphate carboxylase/oxygen- ase genes in Rhodobacter sphaeroides.. Journal of Bacteriology 170:5–11
    [Google Scholar]
  12. Figurski D. 1979; Replication of an origin-containing derivative of the plasmid RK2 dependent on a plasmid function provided in trans. Proceedings of the National Academy of Sciences of the United States of America 76:1648–1652
    [Google Scholar]
  13. Figurski D., Meyer R., Miller D.S., Helinski D.R. 1976; Generation in vitro of deletions in the broad host range plasmid RK2 using phage Mu insertions and a restriction endonuclease. Gene 1:107–119
    [Google Scholar]
  14. Fornari C.S., Kaplan S. 1983; Identification of nitrogenase and carboxylase genes in the photosynthetic bacteria and cloning of a carboxylase gene from Rhodopseudomonas sphaeroides.. Gene 25:291–299
    [Google Scholar]
  15. Fornari C.S., Watkins M., Kaplan S. 1984; Plasmid distribution and analysis in Rhodopseudomonas sphaeroides.. Plasmids 11:39–47
    [Google Scholar]
  16. Gibson J.L., Tabita F.R. 1977a; Different molecular forms of d-ribulose 1,5-bisphosphate carboxylase from Rhodopseudomonas sphaeroides.. Journal of Biological Chemistry 252:943–949
    [Google Scholar]
  17. Gibson J.L., Tabita F.R. 1977b; Characterization of antiserum directed against form II ribulose 1,5-bisphosphate carboxylase from Rhodopseudomonas sphaeroides.. Journal of Bacteriology 131:1020–1022
    [Google Scholar]
  18. Gibson J.L., Tabita F.R. 1977c; Isolation and preliminary characterization of two forms of ribulose 1,5-bisphosphate carboxylase from Rhodopseudomonas capsulata.. Journal of Bacteriology 132:818–823
    [Google Scholar]
  19. Gibson J.L., Tabita F.R. 1986; Isolation of the Rhodopseudomonas sphaeroides form I ribulose 1,5- bisphosphate carboxylase/oxygenase large and small subunit genes and expression of the active hexadeca- meric enzyme in Escherichia coli.. Gene 44:271–278
    [Google Scholar]
  20. Gibson J.L., Tabita F.R. 1987; Organization of phosphoribulokinase and ribulose bisphosphate carboxylase/oxygenase genes in Rhodopseudomonas (Rhodobacter) sphaeroides.. Journal of Bacteriology 169:3685–3690
    [Google Scholar]
  21. Gibson J.L., Tabita F.R. 1988; Localization and mapping of CO2 fixation genes within two gene clusters in Rhodobacter sphaeroides.. Journal of Bacteriology 170:2153–2158
    [Google Scholar]
  22. Glick B.R., Brooks H.E., Pasternak J.J. 1986; Physiological effects of plasmid DNA transformation on Azotobacter vinelandii.. Canadian Journal of Microbiology 32:145–148
    [Google Scholar]
  23. Hallenbeck P., Kaplan S. 1987; Cloning of the gene for phosphoribulokinase activity from Rhodobacter sphaeroides and its expression in Escherichia coli.. Journal of Bacteriology 169:3669–3678
    [Google Scholar]
  24. Ish-Horowitz D., Burke J.F. 1981; Rapid and efficient cosmid vector cloning. Nucleic Acids Research 9:2989–2998
    [Google Scholar]
  25. Jouanneau Y., Tabita F.R. 1986; Independent regulation of synthesis of form I and form II ribulose bisphosphate carboxylase-oxygenase in Rhodopseudomonas sphaeroides.. Journal of Bacteriology 165:620–624
    [Google Scholar]
  26. Knauf V.C., Nester E.W. 1982; Wide host range cloning vectors: a cosmid clone bank of an Agrobacterium Ti plasmid. Plasmid 8:45–54
    [Google Scholar]
  27. Maniatis T., Fritsch E.F., Sambrook J. 1982 Molecular Cloning: a Laboratory Manual. Cold Spring Harbor, NY:: Cold Spring Harbor Laboratory;
    [Google Scholar]
  28. Markwell M.A., Hass S.M., Buber L.L., Tolbert N.E. 1978; A modification of the Lowry procedure to simplify protein determination in membrane and lipoprotein samples. Analytical Biochemistry 87:206–210
    [Google Scholar]
  29. Marrs B. 1981; Mobilization of the genes for photosynthesis from Rhodopseudomonas capsulata by a promiscuous plasmid. Journal of Bacteriology 146:1003–1012
    [Google Scholar]
  30. Miziorko H.M., Lorimer G.H. 1983; Ribulose 1.5-bisphosphate carboxylase-oxygenase. Annual Review of Biochemistry 52:507–535
    [Google Scholar]
  31. Muller E.D., Chory J., Kaplan S. 1985; Cloning and characterization of the gene product of the form II ribulose 1,5-bisphosphate carboxylase gene of Rhodopseudomonas sphaeroides.. Journal of Bacteriology 161:469–472
    [Google Scholar]
  32. Nano F.E., Kaplan S. 1984; Plasmid rearrangements in the photosynthetic bacterium Rhodopseudomonas sphaeroides.. Journal of Bacteriology 158:1094–1103
    [Google Scholar]
  33. Ormerod J.G., Ormerod K.D., Gest H. 1961; Light dependent utilization of organic compounds and photoproduction of hydrogen by photosynthetic bacteria: relationship with nitrogen metabolism. Archives of Biochemistry and Biophysics 94:449–463
    [Google Scholar]
  34. Quivey R.G.Jr Tabita F.R. 1984; Cloning and expression in Escherichia coli of the form II ribulose 1,5 bisphosphate carboxylase/oxygenase gene from Rhodopseudomonas sphaeroides.. Gene 31:91–101
    [Google Scholar]
  35. Rigby P.W.J., Deickmann M., Rhodes C., Berg P. 1977; Labeling deoxyribonucleic acid to high specific activity in vitro by nick translation with DNA polymerase I. Journal of Molecular Biology 113:237–251
    [Google Scholar]
  36. Shively J.M., Davidson T., Marrs B.L. 1984; Derepression in the synthesis of the intermediate and large forms of ribulose 1,5-bisphosphate carboxylase/oxygenase in Rhodopseudomonas capsulata.. Archives of Microbiology 138:233–236
    [Google Scholar]
  37. Siegal R., Ryu D.D.Y. 1985; Kinetic study of instability of recombinant plasmid pPLc23trpAI in E. coli using a two stage continuous culture system. Biotechnology and Bioengineering 27:28–33
    [Google Scholar]
  38. Sistrom W.R. 1977; Transfer of chromosomal genes mediated by plasmid R68.45 in Rhodopseudomonas sphaeroides.. Journal of Bacteriology 131:526–532
    [Google Scholar]
  39. Sistrom W.R., Macaluso, Pledger R. 1984; Mutants of Rhodopseudomonas sphaeroides useful in genetic analysis. Archives of Microbiology 138:161–165
    [Google Scholar]
  40. Stankowski L.R.Jr Taindall K.R., Hsie A.W. 1986; Quantitative and molecular analysis of ethyl methanesulfonate and ICR 191-Induced mutations in AS52 cells. Mutation Research 160:133–147
    [Google Scholar]
  41. Tabita F.R. 1981; Molecular regulation of carbon dioxide assimilation in autotrophic organisms. In Microbial Growth on C1 Compounds, pp 70–81 Dalton H. Edited by London: Heydon & Sons;
    [Google Scholar]
  42. Tabita F.R. 1988; Molecular and cellular regulation of autotrophic carbon dioxide fixation in microorganisms. Microbiological Reviews 52:155–189
    [Google Scholar]
  43. Tabita F.R., Caruso P., Whitman W. 1978; Facile assay of enzymes unique to the Calvin cycle, with special reference to ribulose 1,5-bisphosphate carboxylase. Analytical Biochemistry 84:462–472
    [Google Scholar]
  44. Vieira J., Messing J. 1982; The pUC plasmids, an M13mp7-derived system for insertion mutagenesis and sequencing with synthetic universal primers. Gene 19:259–268
    [Google Scholar]
  45. Weaver K.E. 1985; Molecular regulation of ribulose1,5-bisphosphate carboxylase/oxygenase in Rhodo-pseudomonas sphaeroides. PhD dissertation; University of Texas at Austin, Austin, Texas.:
    [Google Scholar]
  46. Weaver K.E., Tabita F.R. 1983; Isolation and partial characterization of Rhodopseudomonas sphaeroides mutants defective in the regulation of ribulose biphosphate carboxylase/oxygenase. Journal of Bacteriology 156:507–515
    [Google Scholar]
  47. Weaver K.E., Tabita F.R. 1985; Complementation of a Rhodopseudomonas sphaeroides ribulose bisphosphate carboxylase-oxygenase regulatory mutant from a genomic library. Journal of Bacteriology 164:147–154
    [Google Scholar]
  48. Whitman W., Tabita F.R. 1976; Inhibition of ribulose 1,5-bisphosphate carboxylase by pyridoxal 5-phosphate. Biochemical and Biophysical Research Communications 71:1034–1039
    [Google Scholar]
  49. Willison J.C., Vignais P.M. 1982; The use of metronidazole to isolate Nif mutants of Rhodopseudomonas capsulata and the identification of a mutant with altered regulatory properties of nitrogenase. Journal of General Microbiology 128:3001–3010
    [Google Scholar]
  50. Willison J.C., Magnin J.P., Vignais P.M. 1987; Isolation and characterization of Rhodobacter capsulatus strains lacking endogenous plasmids. Archives of Microbiology 147:134–142
    [Google Scholar]
  51. Yanish-Perron C., Vieira J., Messing J. 1985; Improved Ml3 phage cloning vectors and host strains: nucleotide sequences of the M13mp18 and pUC19 vectors. Gene 33:103–119
    [Google Scholar]
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