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. 1995 Feb;177(3):508–516. doi: 10.1128/jb.177.3.508-516.1995

Specific binding of Synechococcus sp. strain PCC 7942 proteins to the enhancer element of psbAII required for high-light-induced expression.

R Li 1, N S Dickerson 1, U W Mueller 1, S S Golden 1
PMCID: PMC176621  PMID: 7836280

Abstract

The psbAII gene of the cyanobacterium Synechococcus sp. strain PCC 7942 is a member of a three-gene family that encodes the D1 protein of the photosystem II reaction center. Transcription of psbAII is rapidly induced when the light intensity reaching the culture increases from 125 microE.m-2.s-1 (low light) to 750 microE.m-2.s-1 (high light). The DNA segment upstream of psbAII that corresponds to the untranslated leader of its major transcript has enhancer activity and confers high-light induction. We show that one or more soluble proteins from PCC 7942 specifically bind to this region of psbAII (designated the enhancer element). In vivo footprinting showed protein binding to the enhancer element in high-light-exposed cell samples but not in those maintained at low light, even though in vitro mobility shifts were detectable with extracts from low- or high-light-grown cells. When 12 bp were deleted from the psbAII enhancer element, protein binding was impaired and high-light induction of both transcriptional and translational psbAII-lacZ reporters was significantly reduced. This finding indicates that protein binding to this region is required for high-light induction of psbAII. The mutant element also showed impaired enhancer activity when combined with a heterologous promoter.

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Selected References

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  1. 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]
  2. Bustos S. A., Schaefer M. R., Golden S. S. Different and rapid responses of four cyanobacterial psbA transcripts to changes in light intensity. J Bacteriol. 1990 Apr;172(4):1998–2004. doi: 10.1128/jb.172.4.1998-2004.1990. [DOI] [PMC free article] [PubMed] [Google Scholar]
  3. Casadaban M. J., Cohen S. N. Analysis of gene control signals by DNA fusion and cloning in Escherichia coli. J Mol Biol. 1980 Apr;138(2):179–207. doi: 10.1016/0022-2836(80)90283-1. [DOI] [PubMed] [Google Scholar]
  4. Casey E. S., Grossman A. In vivo and in vitro characterization of the light-regulated cpcB2A2 promoter of Fremyella diplosiphon. J Bacteriol. 1994 Oct;176(20):6362–6374. doi: 10.1128/jb.176.20.6362-6374.1994. [DOI] [PMC free article] [PubMed] [Google Scholar]
  5. 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]
  6. Chiang G. G., Schaefer M. R., Grossman A. R. Complementation of a red-light-indifferent cyanobacterial mutant. Proc Natl Acad Sci U S A. 1992 Oct 15;89(20):9415–9419. doi: 10.1073/pnas.89.20.9415. [DOI] [PMC free article] [PubMed] [Google Scholar]
  7. Fujiwara S., Zielinski N. A., Chakrabarty A. M. Enhancer-like activity of A1gR1-binding site in alginate gene activation: positional, orientational, and sequence specificity. J Bacteriol. 1993 Sep;175(17):5452–5459. doi: 10.1128/jb.175.17.5452-5459.1993. [DOI] [PMC free article] [PubMed] [Google Scholar]
  8. Gober J. W., Shapiro L. A developmentally regulated Caulobacter flagellar promoter is activated by 3' enhancer and IHF binding elements. Mol Biol Cell. 1992 Aug;3(8):913–926. doi: 10.1091/mbc.3.8.913. [DOI] [PMC free article] [PubMed] [Google Scholar]
  9. 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]
  10. Grossman A. R., Schaefer M. R., Chiang G. G., Collier J. L. The phycobilisome, a light-harvesting complex responsive to environmental conditions. Microbiol Rev. 1993 Sep;57(3):725–749. doi: 10.1128/mr.57.3.725-749.1993. [DOI] [PMC free article] [PubMed] [Google Scholar]
  11. Harter K., Kircher S., Frohnmeyer H., Krenz M., Nagy F., Schäfer E. Light-regulated modification and nuclear translocation of cytosolic G-box binding factors in parsley. Plant Cell. 1994 Apr;6(4):545–559. doi: 10.1105/tpc.6.4.545. [DOI] [PMC free article] [PubMed] [Google Scholar]
  12. Kim J., Zwieb C., Wu C., Adhya S. Bending of DNA by gene-regulatory proteins: construction and use of a DNA bending vector. Gene. 1989 Dec 21;85(1):15–23. doi: 10.1016/0378-1119(89)90459-9. [DOI] [PubMed] [Google Scholar]
  13. Kulkarni R. D., Golden S. S. Adaptation to high light intensity in Synechococcus sp. strain PCC 7942: regulation of three psbA genes and two forms of the D1 protein. J Bacteriol. 1994 Feb;176(4):959–965. doi: 10.1128/jb.176.4.959-965.1994. [DOI] [PMC free article] [PubMed] [Google Scholar]
  14. Kulkarni R. D., Schaefer M. R., Golden S. S. Transcriptional and posttranscriptional components of psbA response to high light intensity in Synechococcus sp. strain PCC 7942. J Bacteriol. 1992 Jun;174(11):3775–3781. doi: 10.1128/jb.174.11.3775-3781.1992. [DOI] [PMC free article] [PubMed] [Google Scholar]
  15. Kunkel T. A., Roberts J. D., Zakour R. A. Rapid and efficient site-specific mutagenesis without phenotypic selection. Methods Enzymol. 1987;154:367–382. doi: 10.1016/0076-6879(87)54085-x. [DOI] [PubMed] [Google Scholar]
  16. Kuwabara M. D., Sigman D. S. Footprinting DNA-protein complexes in situ following gel retardation assays using 1,10-phenanthroline-copper ion: Escherichia coli RNA polymerase-lac promoter complexes. Biochemistry. 1987 Nov 17;26(23):7234–7238. doi: 10.1021/bi00397a006. [DOI] [PubMed] [Google Scholar]
  17. Li R., Golden S. S. Enhancer activity of light-responsive regulatory elements in the untranslated leader regions of cyanobacterial psbA genes. Proc Natl Acad Sci U S A. 1993 Dec 15;90(24):11678–11682. doi: 10.1073/pnas.90.24.11678. [DOI] [PMC free article] [PubMed] [Google Scholar]
  18. Parkinson J. S. Signal transduction schemes of bacteria. Cell. 1993 Jun 4;73(5):857–871. doi: 10.1016/0092-8674(93)90267-t. [DOI] [PubMed] [Google Scholar]
  19. Ramasubramanian T. S., Wei T. F., Golden J. W. Two Anabaena sp. strain PCC 7120 DNA-binding factors interact with vegetative cell- and heterocyst-specific genes. J Bacteriol. 1994 Mar;176(5):1214–1223. doi: 10.1128/jb.176.5.1214-1223.1994. [DOI] [PMC free article] [PubMed] [Google Scholar]
  20. Sasse-Dwight S., Gralla J. D. Footprinting protein-DNA complexes in vivo. Methods Enzymol. 1991;208:146–168. doi: 10.1016/0076-6879(91)08012-7. [DOI] [PubMed] [Google Scholar]
  21. Schaefer M. R., Golden S. S. Differential expression of members of a cyanobacterial psbA gene family in response to light. J Bacteriol. 1989 Jul;171(7):3973–3981. doi: 10.1128/jb.171.7.3973-3981.1989. [DOI] [PMC free article] [PubMed] [Google Scholar]
  22. 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]
  23. Schulze-Lefert P., Dangl J. L., Becker-André M., Hahlbrock K., Schulz W. Inducible in vivo DNA footprints define sequences necessary for UV light activation of the parsley chalcone synthase gene. EMBO J. 1989 Mar;8(3):651–656. doi: 10.1002/j.1460-2075.1989.tb03422.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  24. Shapira S. K., Chou J., Richaud F. V., Casadaban M. J. New versatile plasmid vectors for expression of hybrid proteins coded by a cloned gene fused to lacZ gene sequences encoding an enzymatically active carboxy-terminal portion of beta-galactosidase. Gene. 1983 Nov;25(1):71–82. doi: 10.1016/0378-1119(83)90169-5. [DOI] [PubMed] [Google Scholar]
  25. Sobczyk A., Bely A., Tandeau de Marsac N., Houmard J. A phosphorylated DNA-binding protein is specific for the red-light signal during complementary chromatic adaptation in cyanobacteria. Mol Microbiol. 1994 Sep;13(5):875–885. doi: 10.1111/j.1365-2958.1994.tb00479.x. [DOI] [PubMed] [Google Scholar]
  26. 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]
  27. Stock J. B., Ninfa A. J., Stock A. M. Protein phosphorylation and regulation of adaptive responses in bacteria. Microbiol Rev. 1989 Dec;53(4):450–490. doi: 10.1128/mr.53.4.450-490.1989. [DOI] [PMC free article] [PubMed] [Google Scholar]
  28. Tsinoremas N. F., Schaefer M. R., Golden S. S. Blue and red light reversibly control psbA expression in the cyanobacterium Synechococcus sp. strain PCC 7942. J Biol Chem. 1994 Jun 10;269(23):16143–16147. [PubMed] [Google Scholar]
  29. Weiss D. S., Batut J., Klose K. E., Keener J., Kustu S. The phosphorylated form of the enhancer-binding protein NTRC has an ATPase activity that is essential for activation of transcription. Cell. 1991 Oct 4;67(1):155–167. doi: 10.1016/0092-8674(91)90579-n. [DOI] [PubMed] [Google Scholar]

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