Skip to main content

PMC Search Update

PMC Beta search will replace the current PMC search the week of September 7, 2025. Try out PMC Beta search now and give us your feedback. Learn more

Journal of Bacteriology logoLink to Journal of Bacteriology
. 1994 Mar;176(5):1214–1223. doi: 10.1128/jb.176.5.1214-1223.1994

Two Anabaena sp. strain PCC 7120 DNA-binding factors interact with vegetative cell- and heterocyst-specific genes.

T S Ramasubramanian 1, T F Wei 1, J W Golden 1
PMCID: PMC205182  PMID: 8113160

Abstract

The DNA-binding factor BifA (previously called VF1) binds upstream of the developmentally regulated site-specific recombinase gene xisA in the cyanobacterium Anabaena sp. strain PCC 7120. Besides binding xisA, BifA also binds the glnA, rbcL, and nifH promoter regions. DNase I footprint analysis of BifA binding to glnA showed a protected region -125 to -148 bp upstream of the translation start site. The binding site is between the major glnA transcription start site used in vegetative cells (RNAII) and the major transcription start site used under nitrogen-deficient conditions (RNAI). The two BifA-binding sites on the rbcL promoter were localized to a 24-bp region from +12 to -12 nucleotides and to a 12-bp region from -43 to -54 nucleotides with respect to the transcription start site. Comparison of the BifA binding sites on the glnA, xisA, and rbcL upstream regions revealed the consensus recognition sequence TGT(N9 or 10) ACA. We have identified a second DNA-binding activity (factor 2) that interacts with rbcL and xisA upstream regions. Factor 2 can be resolved from BifA by heparin-Sepharose chromatography and was present in a bifA mutant. Analysis of partially purified vegetative cell and heterocyst extracts showed that whereas BifA was present in both cell types, factor 2 was present only in vegetative cells. DNase I footprint analysis of factor 2 binding to rbcL showed protection of a 63-bp region between positions -15 and -77 with respect to the transcription start site. The factor 2 binding site on xisA was localized to a 68-bp region that showed considerable overlap with the BifA binding sites.

Full text

PDF
1214

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. Batut J., Daveran-Mingot M. L., David M., Jacobs J., Garnerone A. M., Kahn D. fixK, a gene homologous with fnr and crp from Escherichia coli, regulates nitrogen fixation genes both positively and negatively in Rhizobium meliloti. EMBO J. 1989 Apr;8(4):1279–1286. doi: 10.1002/j.1460-2075.1989.tb03502.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  3. Botsford J. L., Harman J. G. Cyclic AMP in prokaryotes. Microbiol Rev. 1992 Mar;56(1):100–122. doi: 10.1128/mr.56.1.100-122.1992. [DOI] [PMC free article] [PubMed] [Google Scholar]
  4. Bradford M. M. A rapid and sensitive method for the quantitation of microgram quantities of protein utilizing the principle of protein-dye binding. Anal Biochem. 1976 May 7;72:248–254. doi: 10.1016/0003-2697(76)90527-3. [DOI] [PubMed] [Google Scholar]
  5. Brahamsha B., Haselkorn R. Identification of multiple RNA polymerase sigma factor homologs in the cyanobacterium Anabaena sp. strain PCC 7120: cloning, expression, and inactivation of the sigB and sigC genes. J Bacteriol. 1992 Nov;174(22):7273–7282. doi: 10.1128/jb.174.22.7273-7282.1992. [DOI] [PMC free article] [PubMed] [Google Scholar]
  6. 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]
  7. 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]
  8. Bustos S. A., Golden S. S. Expression of the psbDII gene in Synechococcus sp. strain PCC 7942 requires sequences downstream of the transcription start site. J Bacteriol. 1991 Dec;173(23):7525–7533. doi: 10.1128/jb.173.23.7525-7533.1991. [DOI] [PMC free article] [PubMed] [Google Scholar]
  9. Buttner M. J. RNA polymerase heterogeneity in Streptomyces coelicolor A3(2). Mol Microbiol. 1989 Nov;3(11):1653–1659. doi: 10.1111/j.1365-2958.1989.tb00151.x. [DOI] [PubMed] [Google Scholar]
  10. 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]
  11. Collado-Vides J., Magasanik B., Gralla J. D. Control site location and transcriptional regulation in Escherichia coli. Microbiol Rev. 1991 Sep;55(3):371–394. doi: 10.1128/mr.55.3.371-394.1991. [DOI] [PMC free article] [PubMed] [Google Scholar]
  12. 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]
  13. Frías J. E., Mérida A., Herrero A., Martín-Nieto J., Flores E. General distribution of the nitrogen control gene ntcA in cyanobacteria. J Bacteriol. 1993 Sep;175(17):5710–5713. doi: 10.1128/jb.175.17.5710-5713.1993. [DOI] [PMC free article] [PubMed] [Google Scholar]
  14. Gaston K., Bell A., Busby S., Fried M. A comparison of the DNA bending activities of the DNA binding proteins CRP and TFIID. Nucleic Acids Res. 1992 Jul 11;20(13):3391–3396. doi: 10.1093/nar/20.13.3391. [DOI] [PMC free article] [PubMed] [Google Scholar]
  15. 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]
  16. 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]
  17. Golden J. W., Whorff L. L., Wiest D. R. Independent regulation of nifHDK operon transcription and DNA rearrangement during heterocyst differentiation in the cyanobacterium Anabaena sp. strain PCC 7120. J Bacteriol. 1991 Nov;173(22):7098–7105. doi: 10.1128/jb.173.22.7098-7105.1991. [DOI] [PMC free article] [PubMed] [Google Scholar]
  18. 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]
  19. Gussin G. N., Ronson C. W., Ausubel F. M. Regulation of nitrogen fixation genes. Annu Rev Genet. 1986;20:567–591. doi: 10.1146/annurev.ge.20.120186.003031. [DOI] [PubMed] [Google Scholar]
  20. 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]
  21. Kraft R., Tardiff J., Krauter K. S., Leinwand L. A. Using mini-prep plasmid DNA for sequencing double stranded templates with Sequenase. Biotechniques. 1988 Jun;6(6):544-6, 549. [PubMed] [Google Scholar]
  22. 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]
  23. Laudenbach D. E., Grossman A. R. Characterization and mutagenesis of sulfur-regulated genes in a cyanobacterium: evidence for function in sulfate transport. J Bacteriol. 1991 May;173(9):2739–2750. doi: 10.1128/jb.173.9.2739-2750.1991. [DOI] [PMC free article] [PubMed] [Google Scholar]
  24. Losick R., Stragier P. Crisscross regulation of cell-type-specific gene expression during development in B. subtilis. Nature. 1992 Feb 13;355(6361):601–604. doi: 10.1038/355601a0. [DOI] [PubMed] [Google Scholar]
  25. 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]
  26. Morett E., Buck M. In vivo studies on the interaction of RNA polymerase-sigma 54 with the Klebsiella pneumoniae and Rhizobium meliloti nifH promoters. The role of NifA in the formation of an open promoter complex. J Mol Biol. 1989 Nov 5;210(1):65–77. doi: 10.1016/0022-2836(89)90291-x. [DOI] [PubMed] [Google Scholar]
  27. Morett E., Fischer H. M., Hennecke H. Influence of oxygen on DNA binding, positive control, and stability of the Bradyrhizobium japonicum NifA regulatory protein. J Bacteriol. 1991 Jun;173(11):3478–3487. doi: 10.1128/jb.173.11.3478-3487.1991. [DOI] [PMC free article] [PubMed] [Google Scholar]
  28. Nierzwicki-Bauer S. A., Curtis S. E., Haselkorn R. Cotranscription of genes encoding the small and large subunits of ribulose-1,5-bisphosphate carboxylase in the cyanobacterium Anabaena 7120. Proc Natl Acad Sci U S A. 1984 Oct;81(19):5961–5965. doi: 10.1073/pnas.81.19.5961. [DOI] [PMC free article] [PubMed] [Google Scholar]
  29. Reznikoff W. S. Catabolite gene activator protein activation of lac transcription. J Bacteriol. 1992 Feb;174(3):655–658. doi: 10.1128/jb.174.3.655-658.1992. [DOI] [PMC free article] [PubMed] [Google Scholar]
  30. Schmidt-Goff C. M., Federspiel N. A. In vivo and in vitro footprinting of a light-regulated promoter in the cyanobacterium Fremyella diplosiphon. J Bacteriol. 1993 Mar;175(6):1806–1813. doi: 10.1128/jb.175.6.1806-1813.1993. [DOI] [PMC free article] [PubMed] [Google Scholar]
  31. Sobczyk A., Schyns G., Tandeau de Marsac N., Houmard J. Transduction of the light signal during complementary chromatic adaptation in the cyanobacterium Calothrix sp. PCC 7601: DNA-binding proteins and modulation by phosphorylation. EMBO J. 1993 Mar;12(3):997–1004. doi: 10.1002/j.1460-2075.1993.tb05740.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  32. Vega-Palas M. A., Flores E., Herrero A. NtcA, a global nitrogen regulator from the cyanobacterium Synechococcus that belongs to the Crp family of bacterial regulators. Mol Microbiol. 1992 Jul;6(13):1853–1859. doi: 10.1111/j.1365-2958.1992.tb01357.x. [DOI] [PubMed] [Google Scholar]
  33. Vega-Palas M. A., Madueño F., Herrero A., Flores E. Identification and cloning of a regulatory gene for nitrogen assimilation in the cyanobacterium Synechococcus sp. strain PCC 7942. J Bacteriol. 1990 Feb;172(2):643–647. doi: 10.1128/jb.172.2.643-647.1990. [DOI] [PMC free article] [PubMed] [Google Scholar]
  34. Wei T. F., Ramasubramanian T. S., Pu F., Golden J. W. Anabaena sp. strain PCC 7120 bifA gene encoding a sequence-specific DNA-binding protein cloned by in vivo transcriptional interference selection. J Bacteriol. 1993 Jul;175(13):4025–4035. doi: 10.1128/jb.175.13.4025-4035.1993. [DOI] [PMC free article] [PubMed] [Google Scholar]

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

RESOURCES