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. 1994 Sep;176(17):5277–5283. doi: 10.1128/jb.176.17.5277-5283.1994

A third genetic locus required for the formation of heterocysts in Anabaena sp. strain PCC 7120.

F Fernández-Piñas 1, F Leganés 1, C P Wolk 1
PMCID: PMC196711  PMID: 8071202

Abstract

Mutagenesis of Anabaena sp. strain PCC 7120 with a derivative of transposon Tn5 led to the isolation of a mutant strain, P6, in which heterocysts are not formed (A. Ernst, T. Black, Y. Cai, J.-M. Panoff, D. N. Tiwari, and C. P. Wolk, J. Bacteriol. 174:6025-6032, 1992). Reconstruction of the transposon mutation of P6 in the wild-type strain reproduced the phenotype of the original mutant. Analysis by pulsed-field gel electrophoresis localized the transposition at ca. 3.44 Mb on the physical map of the chromosome of wild-type Anabaena sp. The transposon was situated within an open reading frame (ORF), which we denote hetP, whose wild-type form was cloned and also sequenced. The predicted HetP protein was not found to show significant sequence similarity to other proteins. The mutation in strain P6 could be complemented by a clone of a fragment of wild-type DNA that includes hetP and at least one additional ORF 3' from hetP, but not by a clone that includes hetP as its only ORF. The latter clone proved highly toxic. The phenotype of the P6 mutant may, therefore, be due to a polar effect of the insertion of the transposon. Filaments of strain P6 and of the wild-type strain, when bearing the complementing fragment on a pDU1-based plasmid, showed an increased frequency of clustered heterocysts compared with that of the wild-type strain.

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Selected References

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  1. Allen M. B., Arnon D. I. Studies on Nitrogen-Fixing Blue-Green Algae. I. Growth and Nitrogen Fixation by Anabaena Cylindrica Lemm. Plant Physiol. 1955 Jul;30(4):366–372. doi: 10.1104/pp.30.4.366. [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. Bancroft I., Wolk C. P. Characterization of an insertion sequence (IS891) of novel structure from the cyanobacterium Anabaena sp. strain M-131. J Bacteriol. 1989 Nov;171(11):5949–5954. doi: 10.1128/jb.171.11.5949-5954.1989. [DOI] [PMC free article] [PubMed] [Google Scholar]
  4. Bancroft I., Wolk C. P., Oren E. V. Physical and genetic maps of the genome of the heterocyst-forming cyanobacterium Anabaena sp. strain PCC 7120. J Bacteriol. 1989 Nov;171(11):5940–5948. doi: 10.1128/jb.171.11.5940-5948.1989. [DOI] [PMC free article] [PubMed] [Google Scholar]
  5. Black T. A., Cai Y., Wolk C. P. Spatial expression and autoregulation of hetR, a gene involved in the control of heterocyst development in Anabaena. Mol Microbiol. 1993 Jul;9(1):77–84. doi: 10.1111/j.1365-2958.1993.tb01670.x. [DOI] [PubMed] [Google Scholar]
  6. Black T. A., Wolk C. P. Analysis of a Het- mutation in Anabaena sp. strain PCC 7120 implicates a secondary metabolite in the regulation of heterocyst spacing. J Bacteriol. 1994 Apr;176(8):2282–2292. doi: 10.1128/jb.176.8.2282-2292.1994. [DOI] [PMC free article] [PubMed] [Google Scholar]
  7. Bradley S., Carr N. G. Heterocyst and nitrogenase development in Anabaena cylindrica. J Gen Microbiol. 1976 Sep;96(1):175–184. doi: 10.1099/00221287-96-1-175. [DOI] [PubMed] [Google Scholar]
  8. Buikema W. J., Haselkorn R. Characterization of a gene controlling heterocyst differentiation in the cyanobacterium Anabaena 7120. Genes Dev. 1991 Feb;5(2):321–330. doi: 10.1101/gad.5.2.321. [DOI] [PubMed] [Google Scholar]
  9. Buikema W. J., Haselkorn R. Isolation and complementation of nitrogen fixation mutants of the cyanobacterium Anabaena sp. strain PCC 7120. J Bacteriol. 1991 Mar;173(6):1879–1885. doi: 10.1128/jb.173.6.1879-1885.1991. [DOI] [PMC free article] [PubMed] [Google Scholar]
  10. Cai Y. P., Wolk C. P. Use of a conditionally lethal gene in Anabaena sp. strain PCC 7120 to select for double recombinants and to entrap insertion sequences. J Bacteriol. 1990 Jun;172(6):3138–3145. doi: 10.1128/jb.172.6.3138-3145.1990. [DOI] [PMC free article] [PubMed] [Google Scholar]
  11. Cohn D. H., Mileham A. J., Simon M. I., Nealson K. H., Rausch S. K., Bonam D., Baldwin T. O. Nucleotide sequence of the luxA gene of Vibrio harveyi and the complete amino acid sequence of the alpha subunit of bacterial luciferase. J Biol Chem. 1985 May 25;260(10):6139–6146. [PubMed] [Google Scholar]
  12. Currier T. C., Haury J. F., Wolk C. P. Isolation and preliminary characterization of auxotrophs of a filamentous Cyanobacterium. J Bacteriol. 1977 Mar;129(3):1556–1562. doi: 10.1128/jb.129.3.1556-1562.1977. [DOI] [PMC free article] [PubMed] [Google Scholar]
  13. Elhai J., Wolk C. P. A versatile class of positive-selection vectors based on the nonviability of palindrome-containing plasmids that allows cloning into long polylinkers. Gene. 1988 Aug 15;68(1):119–138. doi: 10.1016/0378-1119(88)90605-1. [DOI] [PubMed] [Google Scholar]
  14. Elhai J., Wolk C. P. Conjugal transfer of DNA to cyanobacteria. Methods Enzymol. 1988;167:747–754. doi: 10.1016/0076-6879(88)67086-8. [DOI] [PubMed] [Google Scholar]
  15. Elhai J., Wolk C. P. Developmental regulation and spatial pattern of expression of the structural genes for nitrogenase in the cyanobacterium Anabaena. EMBO J. 1990 Oct;9(10):3379–3388. doi: 10.1002/j.1460-2075.1990.tb07539.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  16. Ernst A., Black T., Cai Y., Panoff J. M., Tiwari D. N., Wolk C. P. Synthesis of nitrogenase in mutants of the cyanobacterium Anabaena sp. strain PCC 7120 affected in heterocyst development or metabolism. J Bacteriol. 1992 Oct;174(19):6025–6032. doi: 10.1128/jb.174.19.6025-6032.1992. [DOI] [PMC free article] [PubMed] [Google Scholar]
  17. Holland D., Wolk C. P. Identification and characterization of hetA, a gene that acts early in the process of morphological differentiation of heterocysts. J Bacteriol. 1990 Jun;172(6):3131–3137. doi: 10.1128/jb.172.6.3131-3137.1990. [DOI] [PMC free article] [PubMed] [Google Scholar]
  18. Hu N. T., Thiel T., Giddings T. H., Jr, Wolk C. P. New Anabaena and Nostoc cyanophages from sewage settling ponds. Virology. 1981 Oct 15;114(1):236–246. doi: 10.1016/0042-6822(81)90269-5. [DOI] [PubMed] [Google Scholar]
  19. Johnston T. C., Thompson R. B., Baldwin T. O. Nucleotide sequence of the luxB gene of Vibrio harveyi and the complete amino acid sequence of the beta subunit of bacterial luciferase. J Biol Chem. 1986 Apr 15;261(11):4805–4811. [PubMed] [Google Scholar]
  20. Kuritz T., Ernst A., Black T. A., Wolk C. P. High-resolution mapping of genetic loci of Anabaena PCC 7120 required for photosynthesis and nitrogen fixation. Mol Microbiol. 1993 Apr;8(1):101–110. doi: 10.1111/j.1365-2958.1993.tb01207.x. [DOI] [PubMed] [Google Scholar]
  21. Leganés F., Fernández-Piñas F., Wolk C. P. Two mutations that block heterocyst differentiation have different effects on akinete differentiation in Nostoc ellipsosporum. Mol Microbiol. 1994 May;12(4):679–684. doi: 10.1111/j.1365-2958.1994.tb01055.x. [DOI] [PubMed] [Google Scholar]
  22. Liang J., Scappino L., Haselkorn R. The patA gene product, which contains a region similar to CheY of Escherichia coli, controls heterocyst pattern formation in the cyanobacterium Anabaena 7120. Proc Natl Acad Sci U S A. 1992 Jun 15;89(12):5655–5659. doi: 10.1073/pnas.89.12.5655. [DOI] [PMC free article] [PubMed] [Google Scholar]
  23. Liang J., Scappino L., Haselkorn R. The patB gene product, required for growth of the cyanobacterium Anabaena sp. strain PCC 7120 under nitrogen-limiting conditions, contains ferredoxin and helix-turn-helix domains. J Bacteriol. 1993 Mar;175(6):1697–1704. doi: 10.1128/jb.175.6.1697-1704.1993. [DOI] [PMC free article] [PubMed] [Google Scholar]
  24. Maldener I., Lockau W., Cai Y. P., Wolk C. P. Calcium-dependent protease of the cyanobacterium Anabaena: molecular cloning and expression of the gene in Escherichia coli, sequencing and site-directed mutagenesis. Mol Gen Genet. 1991 Jan;225(1):113–120. doi: 10.1007/BF00282649. [DOI] [PubMed] [Google Scholar]
  25. Vieira J., Messing J. Production of single-stranded plasmid DNA. Methods Enzymol. 1987;153:3–11. doi: 10.1016/0076-6879(87)53044-0. [DOI] [PubMed] [Google Scholar]
  26. Vieira J., Messing J. The pUC plasmids, an M13mp7-derived system for insertion mutagenesis and sequencing with synthetic universal primers. Gene. 1982 Oct;19(3):259–268. doi: 10.1016/0378-1119(82)90015-4. [DOI] [PubMed] [Google Scholar]
  27. Wilcox M., Mitchison G. J., Smith R. J. Mutants of Anabaena cylindrica altered in heterocyst spacing. Arch Microbiol. 1975 May 5;103(3):219–223. doi: 10.1007/BF00436353. [DOI] [PubMed] [Google Scholar]
  28. Wilcox M., Mitchison G. J., Smith R. J. Pattern formation in the blue-green alga Anabaena. II. Controlled proheterocyst regression. J Cell Sci. 1973 Nov;13(3):637–649. doi: 10.1242/jcs.13.3.637. [DOI] [PubMed] [Google Scholar]
  29. Wilcox M., Mitchison G. J., Smith R. J. Pattern formation in the blue-green alga, Anabaena. I. Basic mechanisms. J Cell Sci. 1973 May;12(3):707–723. doi: 10.1242/jcs.12.3.707. [DOI] [PubMed] [Google Scholar]
  30. Wolk C. P., Cai Y., Cardemil L., Flores E., Hohn B., Murry M., Schmetterer G., Schrautemeier B., Wilson R. Isolation and complementation of mutants of Anabaena sp. strain PCC 7120 unable to grow aerobically on dinitrogen. J Bacteriol. 1988 Mar;170(3):1239–1244. doi: 10.1128/jb.170.3.1239-1244.1988. [DOI] [PMC free article] [PubMed] [Google Scholar]
  31. Wolk C. P., Cai Y., Panoff J. M. Use of a transposon with luciferase as a reporter to identify environmentally responsive genes in a cyanobacterium. Proc Natl Acad Sci U S A. 1991 Jun 15;88(12):5355–5359. doi: 10.1073/pnas.88.12.5355. [DOI] [PMC free article] [PubMed] [Google Scholar]
  32. Wolk C. P., Elhai J., Kuritz T., Holland D. Amplified expression of a transcriptional pattern formed during development of Anabaena. Mol Microbiol. 1993 Feb;7(3):441–445. doi: 10.1111/j.1365-2958.1993.tb01135.x. [DOI] [PubMed] [Google Scholar]
  33. Wolk C. P. Physiological basis of the pattern of vegetative growth of a blue-green alga. Proc Natl Acad Sci U S A. 1967 May;57(5):1246–1251. doi: 10.1073/pnas.57.5.1246. [DOI] [PMC free article] [PubMed] [Google Scholar]
  34. Wolk C. P., Quine M. P. Formation of one-dimensional patterns by stochastic processes and by filamentous blue-green algae. Dev Biol. 1975 Oct;46(2):370–382. doi: 10.1016/0012-1606(75)90113-x. [DOI] [PubMed] [Google Scholar]
  35. Wolk C. P., Vonshak A., Kehoe P., Elhai J. Construction of shuttle vectors capable of conjugative transfer from Escherichia coli to nitrogen-fixing filamentous cyanobacteria. Proc Natl Acad Sci U S A. 1984 Mar;81(5):1561–1565. doi: 10.1073/pnas.81.5.1561. [DOI] [PMC free article] [PubMed] [Google Scholar]
  36. Wood N. B., Haselkorn R. Control of phycobiliprotein proteolysis and heterocyst differentiation in Anabaena. J Bacteriol. 1980 Mar;141(3):1375–1385. doi: 10.1128/jb.141.3.1375-1385.1980. [DOI] [PMC free article] [PubMed] [Google Scholar]

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