Skip to main content
Journal of Bacteriology logoLink to Journal of Bacteriology
. 1991 Nov;173(22):7098–7105. doi: 10.1128/jb.173.22.7098-7105.1991

Independent regulation of nifHDK operon transcription and DNA rearrangement during heterocyst differentiation in the cyanobacterium Anabaena sp. strain PCC 7120.

J W Golden 1, L L Whorff 1, D R Wiest 1
PMCID: PMC209215  PMID: 1938911

Abstract

The filamentous cyanobacterium Anabaena sp. strain PCC 7120 expresses the genes required for nitrogen fixation in terminally differentiated cells called heterocysts. The nifHDK operon encodes the nitrogenase polypeptides and is expressed at high levels in heterocysts. During heterocyst differentiation, an 11-kb DNA element is excised from the nifD gene by site-specific recombination. The xisA gene, located on the 11-kb element, is required for the excision of the element. Transcription and DNA rearrangement of the nifHDK operon both occur late during heterocyst differentiation, about 18 to 24 h after induction, suggesting that the regulation of these events might be coupled. We show that heterocyst-specific transcription and DNA rearrangement of the nifHDK operon are independent of one another. Northern (RNA) analysis of the xisA mutant strain DW12-2.2, which cannot excise the nifD 11-kb element or fix nitrogen, showed that the nifH and nifD genes are transcribed on unrearranged chromosomes. The nifK gene was not transcribed in DW12-2.2, indicating that its expression is dependent on the nifH promoter and excision of the 11-kb element from the operon. A 1.68-kb DNA fragment containing the nifH promoter was deleted from the chromosome to produce the mutant strain LW1. LW1 formed heterocysts but did not grow on nitrogen-free medium and showed no transcription through nifD. Southern analysis of LW1 showed normal excision of the 11-kb element from the nifHDK operon, indicating that transcription from the nifH promoter is not required for the developmentally regulated DNA rearrangement.

Full text

PDF
7098

Images in this article

Selected References

These references are in PubMed. This may not be the complete list of references from this article.

  1. Adams D. G., Carr N. G. The developmental biology of heterocyst and akinete formation in cyanobacteria. Crit Rev Microbiol. 1981;9(1):45–100. doi: 10.3109/10408418109104486. [DOI] [PubMed] [Google Scholar]
  2. 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]
  3. Borthakur D., Basche M., Buikema W. J., Borthakur P. B., Haselkorn R. Expression, nucleotide sequence and mutational analysis of two open reading frames in the nif gene region of Anabaena sp. strain PCC7120. Mol Gen Genet. 1990 Apr;221(2):227–234. doi: 10.1007/BF00261725. [DOI] [PubMed] [Google Scholar]
  4. Brahamsha B., Haselkorn R. Isolation and characterization of the gene encoding the principal sigma factor of the vegetative cell RNA polymerase from the cyanobacterium Anabaena sp. strain PCC 7120. J Bacteriol. 1991 Apr;173(8):2442–2450. doi: 10.1128/jb.173.8.2442-2450.1991. [DOI] [PMC free article] [PubMed] [Google Scholar]
  5. Brusca J. S., Chastain C. J., Golden J. W. Expression of the Anabaena sp. strain PCC 7120 xisA gene from a heterologous promoter results in excision of the nifD element. J Bacteriol. 1990 Jul;172(7):3925–3931. doi: 10.1128/jb.172.7.3925-3931.1990. [DOI] [PMC free article] [PubMed] [Google Scholar]
  6. Brusca J. S., Hale M. A., Carrasco C. D., Golden J. W. Excision of an 11-kilobase-pair DNA element from within the nifD gene in anabaena variabilis heterocysts. J Bacteriol. 1989 Aug;171(8):4138–4145. doi: 10.1128/jb.171.8.4138-4145.1989. [DOI] [PMC free article] [PubMed] [Google Scholar]
  7. 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]
  8. Böhme H., Haselkorn R. Molecular cloning and nucleotide sequence analysis of the gene coding for heterocyst ferredoxin from the cyanobacterium Anabaena sp. strain PCC 7120. Mol Gen Genet. 1988 Oct;214(2):278–285. doi: 10.1007/BF00337722. [DOI] [PubMed] [Google Scholar]
  9. Chastain C. J., Brusca J. S., Ramasubramanian T. S., Wei T. F., Golden J. W. A sequence-specific DNA-binding factor (VF1) from Anabaena sp. strain PCC 7120 vegetative cells binds to three adjacent sites in the xisA upstream region. J Bacteriol. 1990 Sep;172(9):5044–5051. doi: 10.1128/jb.172.9.5044-5051.1990. [DOI] [PMC free article] [PubMed] [Google Scholar]
  10. 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]
  11. Fleming H., Haselkorn R. The program of protein synthesis during heterocyst differentiation in nitrogen-fixing blue-green algae. Cell. 1974 Oct;3(2):169–170. doi: 10.1016/0092-8674(74)90121-4. [DOI] [PubMed] [Google Scholar]
  12. Golden G. M., Guzek D. B., Harris R. R., McKie J. E., Potts R. O. Lipid thermotropic transitions in human stratum corneum. J Invest Dermatol. 1986 Mar;86(3):255–259. doi: 10.1111/1523-1747.ep12285373. [DOI] [PubMed] [Google Scholar]
  13. Golden J. W., Carrasco C. D., Mulligan M. E., Schneider G. J., Haselkorn R. Deletion of a 55-kilobase-pair DNA element from the chromosome during heterocyst differentiation of Anabaena sp. strain PCC 7120. J Bacteriol. 1988 Nov;170(11):5034–5041. doi: 10.1128/jb.170.11.5034-5041.1988. [DOI] [PMC free article] [PubMed] [Google Scholar]
  14. Golden J. W., Mulligan M. E., Haselkorn R. Different recombination site specificity of two developmentally regulated genome rearrangements. Nature. 1987 Jun 11;327(6122):526–529. doi: 10.1038/327526a0. [DOI] [PubMed] [Google Scholar]
  15. Golden J. W., Robinson S. J., Haselkorn R. Rearrangement of nitrogen fixation genes during heterocyst differentiation in the cyanobacterium Anabaena. Nature. 1985 Apr 4;314(6010):419–423. doi: 10.1038/314419a0. [DOI] [PubMed] [Google Scholar]
  16. Golden J. W., Wiest D. R. Genome rearrangement and nitrogen fixation in Anabaena blocked by inactivation of xisA gene. Science. 1988 Dec 9;242(4884):1421–1423. doi: 10.1126/science.3144039. [DOI] [PubMed] [Google Scholar]
  17. Golden S. S., Brusslan J., Haselkorn R. Genetic engineering of the cyanobacterial chromosome. Methods Enzymol. 1987;153:215–231. doi: 10.1016/0076-6879(87)53055-5. [DOI] [PubMed] [Google Scholar]
  18. Haselkorn R., Rice D., Curtis S. E., Robinson S. J. Organization and transcription of genes important in Anabaena heterocyst differentiation. Ann Microbiol (Paris) 1983 Jul-Aug;134B(1):181–193. doi: 10.1016/s0769-2609(83)80104-5. [DOI] [PubMed] [Google Scholar]
  19. Herrero A., Wolk C. P. Genetic mapping of the chromosome of the cyanobacterium, Anabaena variabilis. Proximity of the structural genes for nitrogenase and ribulose-bisphosphate carboxylase. J Biol Chem. 1986 Jun 15;261(17):7748–7754. [PubMed] [Google Scholar]
  20. 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]
  21. Lammers P. J., Golden J. W., Haselkorn R. Identification and sequence of a gene required for a developmentally regulated DNA excision in Anabaena. Cell. 1986 Mar 28;44(6):905–911. doi: 10.1016/0092-8674(86)90013-9. [DOI] [PubMed] [Google Scholar]
  22. Lammers P. J., McLaughlin S., Papin S., Trujillo-Provencio C., Ryncarz A. J., 2nd Developmental rearrangement of cyanobacterial nif genes: nucleotide sequence, open reading frames, and cytochrome P-450 homology of the Anabaena sp. strain PCC 7120 nifD element. J Bacteriol. 1990 Dec;172(12):6981–6990. doi: 10.1128/jb.172.12.6981-6990.1990. [DOI] [PMC free article] [PubMed] [Google Scholar]
  23. Lynn M. E., Bantle J. A., Ownby J. D. Estimation of gene expression in heterocysts of Anabaena variabilis by using DNA-RNA hybridization. J Bacteriol. 1986 Sep;167(3):940–946. doi: 10.1128/jb.167.3.940-946.1986. [DOI] [PMC free article] [PubMed] [Google Scholar]
  24. Meeks J. C., Joseph C. M., Haselkorn R. Organization of the nif genes in cyanobacteria in symbiotic association with Azolla and Anthoceros. Arch Microbiol. 1988 May;150(1):61–71. doi: 10.1007/BF00409719. [DOI] [PubMed] [Google Scholar]
  25. Mulligan M. E., Buikema W. J., Haselkorn R. Bacterial-type ferredoxin genes in the nitrogen fixation regions of the cyanobacterium Anabaena sp. strain PCC 7120 and Rhizobium meliloti. J Bacteriol. 1988 Sep;170(9):4406–4410. doi: 10.1128/jb.170.9.4406-4410.1988. [DOI] [PMC free article] [PubMed] [Google Scholar]
  26. Mulligan M. E., Haselkorn R. Nitrogen fixation (nif) genes of the cyanobacterium Anabaena species strain PCC 7120. The nifB-fdxN-nifS-nifU operon. J Biol Chem. 1989 Nov 15;264(32):19200–19207. [PubMed] [Google Scholar]
  27. Rice D., Mazur B. J., Haselkorn R. Isolation and physical mapping of nitrogen fixation genes from the cyanobacterium Anabaena 7120. J Biol Chem. 1982 Nov 10;257(21):13157–13163. [PubMed] [Google Scholar]
  28. Stragier P., Losick R. Cascades of sigma factors revisited. Mol Microbiol. 1990 Nov;4(11):1801–1806. doi: 10.1111/j.1365-2958.1990.tb02028.x. [DOI] [PubMed] [Google Scholar]
  29. 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]

Articles from Journal of Bacteriology are provided here courtesy of American Society for Microbiology (ASM)

RESOURCES