Mutagenesis of the tal gene-encoding Transaldolase in the Cyanobacterium, Anabaena sp. PCC7120

The transaldolase gene (tal) of Anabaena sp. PCC7120 was interrupted by the insertion of the interposon W. Transaldolase assays showed that the tal mutant strain possessed the same activity as the wild-type, indicating that the second copy of the gene complements the enzyme activity. Being coded by a gene (zwf) downstream of tal and probably in the same operon, glucose-6-phosphate dehydrogenase (G6PDH) activity was also analysed. Only 34% of wild-type G6PDH activity was retained in the tal mutant strain. This may have due to the polar affect of tal mutation on the transcription of the zwf gene. Growth of the tal mutant was not different than that of the wild-type in the presence of combined nitrogen, but the mutant reached the stationary phase faster than the wild-type in the absence of combined nitrogen. This was probably because of the reduction of G6PDH activity, resulting in less production of reductant and energy in heterocysts, which negatively affects nitrogen fixation and growth.

Mutagenesis of the tal gene-encoding Transaldolase in the Cyanobacterium, Anabaena sp. PCC7120

The transaldolase gene (tal) of Anabaena sp. PCC7120 was interrupted by the insertion of the interposon W. Transaldolase assays showed that the tal mutant strain possessed the same activity as the wild-type, indicating that the second copy of the gene complements the enzyme activity. Being coded by a gene (zwf) downstream of tal and probably in the same operon, glucose-6-phosphate dehydrogenase (G6PDH) activity was also analysed. Only 34% of wild-type G6PDH activity was retained in the tal mutant strain. This may have due to the polar affect of tal mutation on the transcription of the zwf gene. Growth of the tal mutant was not different than that of the wild-type in the presence of combined nitrogen, but the mutant reached the stationary phase faster than the wild-type in the absence of combined nitrogen. This was probably because of the reduction of G6PDH activity, resulting in less production of reductant and energy in heterocysts, which negatively affects nitrogen fixation and growth.

___

  • 1. Smith AJ. Modes of cyanobacterial carbon metabolism. In: Carr NG, Whitton BA. ed. The Biology of Cyanobacteria. Blackwell Scientific; 1982: pp. 47-85.
  • 2. Udvardy J, Borbely G, Juhasz A et al. Thioredoxins and the redox modulation of glucose-6-phosphate dehydrogenase in Anabaena sp. strain PCC7120 vegetative cells and heterocysts. J. Bacteriol. 157: 681-683, 1984.
  • 3. Wolk CP. (1982). Heterocysts. In: Carr NG, Whitton BA. ed. The Biology of Cyanobacteria. Blackwell Scientific; 1982: pp. 359- 386.
  • 4. Stanier RY, Cohen-Bazire G. Phototrophic prokaryotes: the cyanobacteria. Ann. Rev. Microbiol. 31: 225-274, 1977.
  • 5. Wolk CP, Ernst A, Elhai J. Heterocyst metabolism and development. In: Bryant DA. ed. The Molecular Biology of Cyanobacteria. Kluwer Academic; 1994: pp. 769-823.
  • 6. Fay P. Oxygen relations of nitrogen fixation in cyanobacteria. Microbiological Reviews 56:340-373, 1992.
  • 7. Schörken U, Thorell S, Schürmann M et al. Identification of catalytically important residues in active site of Escherichia coli transaldolase. Eur. J. Biochem. 268: 2408-2415, 2001.
  • 8. Latzko E, Gibbs M. Enzyme activities of the carbon reduction cycle in some photosynthetic organisms. Plant Physiol. 44: 295-300, 1969.
  • 9. Newman J, Karakaya H, Scanlan DJ et al. A comparison of gene organisation in the zwf region of the genomes of the cyanobacteria Synechococcus sp. PCC7942 and Anabaena sp. PCC7120. FEMS Microbiol. Lett. 133:187-193, 1995.
  • 10. Summers, ML, Meeks JC, Chu S et al. Nucleotide sequence of an operon in Nostoc sp. strain ATCC29133 encoding four genes of the oxidative pentose phosphate cycle. Plant Physiol. 107: 267- 268, 1995a.
  • 11. Kaneko T, Sato S, Kotani H et al. Sequence analysis of the genome of the unicellular cyanobacterium Synechocystis sp. strain PCC6803. II. Sequence determination of the entire genome and assignment of potential protein-coding regions. DNA Res. 3(3), 109-136, 1996.
  • 12. Kaneko T, Nakamura Y, Wolk CP et al. Complete genomic sequence of the filamentous nitrogen-fixing cyanobacterium Anabaena sp. strain PCC7120. DNA Res. 8(5): 205-213, 2001.
  • 13. Summers ML, Meeks JC. Transcriptional regulation of zwf encoding, glucose-6-phosphate dehydrogenase, from the cyanobacterium Nostoc punctiforme strain ATCC 29133. Mol. Microbiol. 22(3), 473-480, 1996.
  • 14. Rippka R, Deruells J, Waterburry JB et al. Generic assignments, strain histories and properties of pure cultures of cyanobacteria. J. Gen. Microbiol. 111: 1-61, 1979.
  • 15. Maniatis T, Fritsch EF, Sambrook J. Molecular cloning: a laboratory manual. Cold Spring Harbor Laboratory, Cold Spring Harbor. New York; 1982.
  • 16. Wertman K, Wyman AR, Botstein D. Host/vector interactions which affect the ability of recombinant phage lambda clones. Gene 49:253-262, 1986.
  • 17. Ward ES, Howe CJ. Troubleshooting in chain-termination DNA sequencing. In: Howe CJ, Ward ES. ed. Nucleic Acids Sequencing. A Practical Approach. IRL Press; 1989: pp 79-97.
  • 18. Prentki P, Karch F, Iida S et al. The plasmid cloning vector pBR325 contains a 482 base-pair-long inverted duplication. Gene 14: 289-299, 1981.
  • 19. Black TA, Cai Y, Wolk CP. Spatial expression and autoregulation of hetR, a gene involved in the control of heterocyst development in Anabaena. Mol. Microbiol. 9: 77-84, 1993.
  • 20. Elhai J, Wolk, CP. Conjugal transfer of DNA to cyanobacteria. In: Parker L, Glazer AN. ed. Methods in Enzymology 167. Academic Press; 1988: 747-754.
  • 21. Thomas CM, Smith CA. Incompatibility group P plasmids: Genetics, evolution, and use in genetic manipulation. Ann. Rev. Microbiol. 41:77-101,1987.
  • 22. Prentki P, Krisch HM. In vitro insertional mutagenesis with a selectable DNA fragment. Gene 29: 303-313, 1984.
  • 23. Cai Y, Wolk CP. Use of conditionally lethal gene in Anabaena sp. strain PCC7120 to select for double recombinants and entrap insertion sequences. J. Bacteriol. 172: 3138-3145, 1990.
  • 24. Scanlan DJ, Sundaram S, Newman J et al. Characterisation of a zwf mutant of Synechococcus sp. strain PCC7942. J. Bacteriol. 177: 2550-2553, 1995.
  • 25. Schaeffer F, Stanier RY. Glucose-6-phosphate dehydrogenase of Anabaena sp. Kinetic and molecular properties. Arch. Microbiol. 116: 9-19, 1978.
  • 26. Levering PR, Dijkhuizen L. Transaldolase isoenzymes from Arthrobacter P1. In: Lidstrom MF. ed. Methods in Enzymology 188. Academic Press; 1990: pp. 405-411.
  • 27. Sprenger GA, Schorken U, Sprenger G et al. Transaldolase B of Escherichia coli K-12: Cloning of its gene, talB, and characterisation of the enzyme from recombinant strains. J. Bacteriol. 177:5930-5936, 1995.
  • 28. Yura T, Mori H, Nagai H et al. Systematic sequencing of the Escherichia coli genome: analysis of the 0-2.4 region. Nucleic Acids Res. 20: 3305-3308, 1992.
  • 29. Schaaff I, Hohmann S Zimmermann FK. Molecular analysis of the structural gene for yeast transaldolase. Eur. J. Biochem. 188: 597-603, 1990.
  • 30. Prentki P, Binda A, Epstein A. Plasmid vectors for selecting IS1- promoted deletions in cloned DNA: sequence analysis of omega interposon. Gene 103: 17-23, 1991.
  • 31. Wolk CP, Vonshak A, Kehoe P et al. Construction of shuttle vectors capable of conjugative transfer from Escherichia coli to nitrogen-fixing filamentous cyanobacteria. Prot. Natl. Acad. Sci. USA 81: 1561-1565, 1984.
  • 32. Moser DP, Zarko D, Kallas T. Characterisation of a restriction barrier and electrotransformation of the cyanobacterium Nostoc PCC 7121. Arch. Microbiol. 160:229-237, 1993.
  • 33. Thiel T, Poo H. Transformation of a filamentous cyanobacterium by electroporation. J. Bacteriol. 171:5743-5746, 1989.
  • 34. Thiel T. Genetic analysis of cyanobacteria. In: Bryant DA. ed. The Molecular Biology of Cyanobacteria. Kluwer Academic; 1994: pp. 1-25.
  • 35. Gay P, le Coq D, Steinmetz M et al. Cloning structural gene sacB, which codes for exoenzyme levansucrase of Bacillus subtilis: expression of the gene in Escherichia coli. J. Bacteriol. 153: 1424-1431, 1983.
  • 36. Summers ML, Wallis JG, Campbell EL et al. Genetic evidence of a major role for glucose-6-phosphate dehydrogenase in nitrogen fixation and dark growth of the cyanobacterium Nostoc sp. strain ATCC29133. J. Bacteriol. 177: 6184-6194, 1995b.
Turkish Journal of Biology-Cover
  • ISSN: 1300-0152
  • Yayın Aralığı: Yılda 6 Sayı
  • Yayıncı: TÜBİTAK
Sayıdaki Diğer Makaleler

Plant Regeneration from Callus Culture of Clematis gouriana Roxb. – A Rare Medicinal Plant

Hanumanaika Raja NAIKA, Venkatarangaiah KRISHNA

Soil Salinity Alters Growth, Chlorophyll Content, and Secondary Metabolite Accumulation in Catharanthus roseus

Cheruth Abdul JALEEL, Beemarao SANKAR, Ramalingam SRIDHARAN, Rajaram PANNEERSELVAM

Improvement of the Nutritional Value of Soybean [Glycine max (L) Merr.] Seed with Alteration in Protein Subunits of Glycinin (11S Globulin) and b-conglycinin (7S Globulin)

Behzad KAVIANI, Ardashir KHARABIAN

A Comparative Study on Colchicine Application Methods in Obtaining Doubled Haploids of Tobacco (Nicotiana tabacum L.)

Betül BÜRÜN, Ülkü EMİROĞLU

Plant regeneration from callus culture of clematis gouriana roxb. – A rare medicinal plant

Hanumanaika RAJA NAIKA, Venkatarangaiah KRISHNA

Evaluation of Antioxidant and Antimicrobial Activity of Seaweed (Sargassum sp.) Extract: A Study on Inhibition of Glutathione-S-Transferase Activity

Jayanta Kumar PATRA, Sakti Kanta RATH, Karmabeer JENA

Cropping Effects on Microbial Population and Nitrogenase Activity in Saline Arid Soil

Dilfuza EGAMBERDIEVA, Zulfiya KUCHAROVA

Mutagenesis of the tal gene-encoding Transaldolase in the Cyanobacterium, Anabaena sp. PCC7120

Haydar KARAKAYA, Nicholas H. MANN

Dispersal Ability and Parasitization Performance of Egg Parasitoid Trichogramma evanescens Westwood (Hymenoptera: Trichogrammatidae) in Field and Storage Conditions

Abdurrahman AYVAZ, Eyüp KARASU, Salih KARABÖRKLÜ, Semih YILMAZ

Plant Regeneration from Callus Culture of Clematis gouriana Roxb. – A Rare Medicinal Plant

Hanumanaika Raja NAIKA, Venkatarangaiah KRISHNA