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. 1988 Jul;170(7):2961–2970. doi: 10.1128/jb.170.7.2961-2970.1988

Transcriptional organization of the phycocyanin subunit gene clusters of the cyanobacterium Anacystis nidulans UTEX 625.

S R Kalla 1, L K Lind 1, J Lidholm 1, P Gustafsson 1
PMCID: PMC211235  PMID: 2454910

Abstract

The phycocyanin subunit gene cluster is duplicated on the chromosome of the cyanobacterium Anacystis nidulans UTEX 625. The two gene clusters cpcB1A1 (left) and cpcB2A2 (right) are separated by about 2,500 base pairs, and in each cluster the beta-subunit gene is located upstream from the alpha-subunit gene. Filter hybridizations with phycocyanin-specific probes to total RNA detected at least two major transcripts that were 1,300 to 1,400 nucleotides long. Besides these major mRNA species, two minor transcripts of 3,400 and 3,700 nucleotides covering one of the gene clusters and the region between the clusters were found. No additional minor transcripts were found in the intergenic region between the two phycocyanin gene clusters. The lengths of the major mRNAs indicated that the beta- and alpha-subunit genes were cotranscribed. No apparent homologies were found when the DNA sequences located upstream from the proposed ribosome-binding site of the two phycocyanin beta-subunit genes were compared. Northern hybridizations with gene cluster-specific probes from the regions 5' of the beta-subunit genes, as well as S1 nuclease mapping and mRNA primer extension experiments, showed that both gene clusters were transcribed. The minor transcripts were found to initiate upstream from the left gene cluster. Two mRNA 5' ends were mapped upstream from the cpcB1A1 gene cluster, while only one 5' end was mapped in front of the cpcB2A2 gene cluster. All transcripts were present in RNA preparations from cultures grown under high levels of white light as well as under low levels of red light. The level of phycocyanin-specific mRNA, measured as part of the total RNA, was lower under low levels of red light compared with that under high levels of white light. Conserved sequence motifs were found when the promoter region of the cpcB1A1 gene cluster and promoter regions from other cyanobacterial photosynthesis genes were compared. The DNA sequences covering the proposed transcriptional attenuators and transcriptional stop signals contained several potential hairpin structures. One potential hairpin structure was located immediately downstream of the left phycocyanin gene cluster and was concluded to limit the level of transcription for the minor transcripts initiating upstream of the cpcB1A1 gene cluster.

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Selected References

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  1. Belasco J. G., Nilsson G., von Gabain A., Cohen S. N. The stability of E. coli gene transcripts is dependent on determinants localized to specific mRNA segments. Cell. 1986 Jul 18;46(2):245–251. doi: 10.1016/0092-8674(86)90741-5. [DOI] [PubMed] [Google Scholar]
  2. Belknap W. R., Haselkorn R. Cloning and light regulation of expression of the phycocyanin operon of the cyanobacterium Anabaena. EMBO J. 1987 Apr;6(4):871–884. doi: 10.1002/j.1460-2075.1987.tb04833.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  3. Conley P. B., Lemaux P. G., Grossman A. R. Cyanobacterial light-harvesting complex subunits encoded in two red light-induced transcripts. Science. 1985 Nov 1;230(4725):550–553. doi: 10.1126/science.3931221. [DOI] [PubMed] [Google Scholar]
  4. Conley P. B., Lemaux P. G., Lomax T. L., Grossman A. R. Genes encoding major light-harvesting polypeptides are clustered on the genome of the cyanobacterium Fremyella diplosiphon. Proc Natl Acad Sci U S A. 1986 Jun;83(11):3924–3928. doi: 10.1073/pnas.83.11.3924. [DOI] [PMC free article] [PubMed] [Google Scholar]
  5. Feinberg A. P., Vogelstein B. A technique for radiolabeling DNA restriction endonuclease fragments to high specific activity. Anal Biochem. 1983 Jul 1;132(1):6–13. doi: 10.1016/0003-2697(83)90418-9. [DOI] [PubMed] [Google Scholar]
  6. Füglistaller P., Suter F., Zuber H. Linker polypeptides of the phycobilisome from the cyanobacterium Mastigocladus laminosus: amino-acid sequences and relationships. Biol Chem Hoppe Seyler. 1985 Oct;366(10):993–1001. doi: 10.1515/bchm3.1985.366.2.993. [DOI] [PubMed] [Google Scholar]
  7. Glazer A. N. Comparative biochemistry of photosynthetic light-harvesting systems. Annu Rev Biochem. 1983;52:125–157. doi: 10.1146/annurev.bi.52.070183.001013. [DOI] [PubMed] [Google Scholar]
  8. Glazer A. N. Phycobilisomes: structure and dynamics. Annu Rev Microbiol. 1982;36:173–198. doi: 10.1146/annurev.mi.36.100182.001133. [DOI] [PubMed] [Google Scholar]
  9. Hamlyn P. H., Gait M. J., Milstein C. Complete sequence of an immunoglobulin mRNA using specific priming and the dideoxynucleotide method of RNA sequencing. Nucleic Acids Res. 1981 Sep 25;9(18):4485–4494. doi: 10.1093/nar/9.18.4485. [DOI] [PMC free article] [PubMed] [Google Scholar]
  10. Harley C. B., Reynolds R. P. Analysis of E. coli promoter sequences. Nucleic Acids Res. 1987 Mar 11;15(5):2343–2361. doi: 10.1093/nar/15.5.2343. [DOI] [PMC free article] [PubMed] [Google Scholar]
  11. Harr R., Fällman P., Häggström M., Wahlström L., Gustafsson P. GENEUS, a computer system for DNA and protein sequence analysis containing an information retrieval system for the EMBL data library. Nucleic Acids Res. 1986 Jan 10;14(1):273–284. doi: 10.1093/nar/14.1.273. [DOI] [PMC free article] [PubMed] [Google Scholar]
  12. Harr R., Häggström M., Gustafsson P. Search algorithm for pattern match analysis of nucleic acid sequences. Nucleic Acids Res. 1983 May 11;11(9):2943–2957. doi: 10.1093/nar/11.9.2943. [DOI] [PMC free article] [PubMed] [Google Scholar]
  13. Hu N., Messing J. The making of strand-specific M13 probes. Gene. 1982 Mar;17(3):271–277. doi: 10.1016/0378-1119(82)90143-3. [DOI] [PubMed] [Google Scholar]
  14. Lau P. C., Condie J. A., Alvarado-Urbina G., Lau R. H. Nucleotide sequence of phycocyanin beta-subunit gene of cyanobacterium Anacystis nidulans strain R2. Nucleic Acids Res. 1987 Mar 11;15(5):2394–2394. doi: 10.1093/nar/15.5.2394. [DOI] [PMC free article] [PubMed] [Google Scholar]
  15. Lau R. H., Alvarado-Urbina G., Lau P. C. Phycocyanin alpha-subunit gene of Anacystis nidulans R2: cloning, nucleotide sequencing and expression in Escherichia coli. Gene. 1987;52(1):21–29. doi: 10.1016/0378-1119(87)90391-x. [DOI] [PubMed] [Google Scholar]
  16. Lemaux P. G., Grossman A. R. Major light-harvesting polypeptides encoded in polycistronic transcripts in a eukaryotic alga. EMBO J. 1985 Aug;4(8):1911–1919. doi: 10.1002/j.1460-2075.1985.tb03870.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  17. Lemaux P. G., Grossman A. Isolation and characterization of a gene for a major light-harvesting polypeptide from Cyanophora paradoxa. Proc Natl Acad Sci U S A. 1984 Jul;81(13):4100–4104. doi: 10.1073/pnas.81.13.4100. [DOI] [PMC free article] [PubMed] [Google Scholar]
  18. Lomax T. L., Conley P. B., Schilling J., Grossman A. R. Isolation and characterization of light-regulated phycobilisome linker polypeptide genes and their transcription as a polycistronic mRNA. J Bacteriol. 1987 Jun;169(6):2675–2684. doi: 10.1128/jb.169.6.2675-2684.1987. [DOI] [PMC free article] [PubMed] [Google Scholar]
  19. Lönneborg A., Lind L. K., Kalla S. R., Gustafsson P., Oquist G. Acclimation Processes in the Light-Harvesting System of the Cyanobacterium Anacystis nidulans following a Light Shift from White to Red Light. Plant Physiol. 1985 May;78(1):110–114. doi: 10.1104/pp.78.1.110. [DOI] [PMC free article] [PubMed] [Google Scholar]
  20. Messing J., Vieira J. A new pair of M13 vectors for selecting either DNA strand of double-digest restriction fragments. Gene. 1982 Oct;19(3):269–276. doi: 10.1016/0378-1119(82)90016-6. [DOI] [PubMed] [Google Scholar]
  21. Mizuno T., Chou M. Y., Inouye M. A unique mechanism regulating gene expression: translational inhibition by a complementary RNA transcript (micRNA). Proc Natl Acad Sci U S A. 1984 Apr;81(7):1966–1970. doi: 10.1073/pnas.81.7.1966. [DOI] [PMC free article] [PubMed] [Google Scholar]
  22. 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]
  23. Nomura M., Gourse R., Baughman G. Regulation of the synthesis of ribosomes and ribosomal components. Annu Rev Biochem. 1984;53:75–117. doi: 10.1146/annurev.bi.53.070184.000451. [DOI] [PubMed] [Google Scholar]
  24. Pilot T. J., Fox J. L. Cloning and sequencing of the genes encoding the alpha and beta subunits of C-phycocyanin from the cyanobacterium Agmenellum quadruplicatum. Proc Natl Acad Sci U S A. 1984 Nov;81(22):6983–6987. doi: 10.1073/pnas.81.22.6983. [DOI] [PMC free article] [PubMed] [Google Scholar]
  25. Rave N., Crkvenjakov R., Boedtker H. Identification of procollagen mRNAs transferred to diazobenzyloxymethyl paper from formaldehyde agarose gels. Nucleic Acids Res. 1979 Aug 10;6(11):3559–3567. doi: 10.1093/nar/6.11.3559. [DOI] [PMC free article] [PubMed] [Google Scholar]
  26. SCHERRER K., DARNELL J. E. Sedimentation characteristics of rapidly labelled RNA from HeLa cells. Biochem Biophys Res Commun. 1962 Jun 4;7:486–490. doi: 10.1016/0006-291x(62)90341-8. [DOI] [PubMed] [Google Scholar]
  27. Stanier R. Y., Cohen-Bazire G. Phototrophic prokaryotes: the cyanobacteria. Annu Rev Microbiol. 1977;31:225–274. doi: 10.1146/annurev.mi.31.100177.001301. [DOI] [PubMed] [Google Scholar]
  28. Tinoco I., Jr, Borer P. N., Dengler B., Levin M. D., Uhlenbeck O. C., Crothers D. M., Bralla J. Improved estimation of secondary structure in ribonucleic acids. Nat New Biol. 1973 Nov 14;246(150):40–41. doi: 10.1038/newbio246040a0. [DOI] [PubMed] [Google Scholar]
  29. 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]
  30. Yanofsky C. Attenuation in the control of expression of bacterial operons. Nature. 1981 Feb 26;289(5800):751–758. doi: 10.1038/289751a0. [DOI] [PubMed] [Google Scholar]
  31. de Lorimier R., Bryant D. A., Porter R. D., Liu W. Y., Jay E., Stevens S. E., Jr Genes for the alpha and beta subunits of phycocyanin. Proc Natl Acad Sci U S A. 1984 Dec;81(24):7946–7950. doi: 10.1073/pnas.81.24.7946. [DOI] [PMC free article] [PubMed] [Google Scholar]

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