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Expression of psbA genes produces prominent 5′ psbA mRNA fragments in Synechococcus sp. PCC 7942

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Abstract

Expression of the psbA genes, which in the cyanobacterium Synechococcus sp. PCC 7942 encode two different forms of the reaction centre D1 protein of photosystem II (D1:1 and D1:2), was studied under different light and temperature conditions. In addition to the mature 1200 nt psbA messages, three shorter mRNA fragments of 220, 320 and 900 nt were also found. All three mRNA fragments could be recognized by using different gene probes from the coding region of the psbAI gene, whereas the corresponding psbAII/III gene probes recognized only the 220 nt mRNA fragment. The 5' 320 nt mRNA fragment from the psbAI gene probably represents a degradation product, since the corresponding 3' 900 nt psbAI mRNA fragment was also detected. By contrast, the 5' 220 nt mRNA fragment of all psbA messages is suggested to be a truncated psbA transcript, since no corresponding 3' fragment was ever found. Inhibition of translation either by a protein synthesis inhibitor or by a shift of cells to lower temperature, increased the number of 1200 nt psbAII/III messages but the number of 5' 220 nt psbAII/III mRNA fragment increased even more dramatically. The first 66 bp after ATG, where the psbAI and psbAII/III genes mostly differ from each other, also appeared important in determining the amount of produced truncated psbA transcripts, as evidenced by the expression of different tac-psbA constructs in the presence of protein synthesis inhibitor. We suggest that both the psbAI and the psbAII/III genes have a latent intragenic termination site and truncated psbA transcripts are produced at high levels under stress conditions when transcription becomes uncoupled from translation.This is to prevent wasting metabolic energy in the production of unused transcripts.

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References

  1. Alifano P, Rivellini F, Limauro D, Bruni CB, Carlomagno MS: A consensus motif common to all Rho-dependent prokaryotic transcription terminators. Cell 64: 553–563 (1991).

    PubMed  Google Scholar 

  2. Arnon DI: Copper enzymes in isolated chloroplasts. Polyphenoloxidase in Beta vulgaris. Plant Physiol 24: 1–15 (1949).

    Google Scholar 

  3. Aymerich S, Steinmetz M: Specificity determinants and structural features in the RNA target of the bacterial antiterminator proteins of the BglG/SacY family. Proc Natl Acad Sci USA 80: 10410–10414 (1992).

    Google Scholar 

  4. Babitzke P, Bear DG, Yanofsky: TRAP, the RNA-binding attenuation protein of Bacillus subtilis, is a toroid shape molecule that binds transcripts containing GAG or UAG repeats separated by two nucleotides. Proc Natl Acad Sci USA 92: 7916–7920 (1995).

    PubMed  Google Scholar 

  5. Bradford MM: A rapid sensitive method for the quantification of microgram quantities of protein using the principle of protein-dye binding. Anal Biochem 72: 248–258 (1976).

    Article  PubMed  Google Scholar 

  6. Burns MC, Richardson JP: NusG is required to overcome a kinetic limitation to Rho function at an intragenic terminator. Proc Natl Acad Sci USA 92: 4732–4742 (1995).

    Google Scholar 

  7. Campbell D, Zhou G, Gustafsson P, Öquist G, Clarke AK: Electron transport regulates exchange of two forms of photosystem II D1 protein in the cyanobacterium Synechococcus. EMBO J 14: 5457–5466 (1995).

    PubMed  Google Scholar 

  8. Clarke AK, Campbell D, Gustafsson P, Öquist G: Dynamic responses of photosystem II and phycobilisomes to changing light in the cyanobacterium Synechococcus sp. PCC 7942. Planta 197: 553–562 (1995).

    Google Scholar 

  9. Fido RJ, Tatham AS, Shewry PR: Application of protein blotting in plant biochemistry and molecular biology. Meth Plant Biochem 10: 101–114 (1993).

    Google Scholar 

  10. Golden SS, Brusslan J, Haselkorn R: Expression of a family of psbA genes encoding photosystem II polypeptide in the cyanobacterium Anacystis nidulans R2. EMBO J 5: 2789–2798 (1986).

    PubMed  Google Scholar 

  11. Green PJ, Kay SA, Lam E, Zhua N-H: In vitro DNA footprinting. In: Gelvin JB, Schilperoort RA (eds) Plant Molecular Biol Manual, pp. B11: 1–22. Kluwer Academic Publishers, Dordrecht, Netherlands (1989).

    Google Scholar 

  12. Henkin TM: Control of transcription termination in procaryotes. Annu Rev Genet 30: 35–57 (1996).

    Google Scholar 

  13. Kulkarni RD, Golden SS: mRNA stability is regulated by a coding-region element and the unique 50 untranslated leader sequences of the three Synechococcus psbA transcripts. Mol Microbiol 24: 1131–1142 (1997).

    PubMed  Google Scholar 

  14. Kulkarni RD, Golden SS: 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 176: 959–965 (1994).

    PubMed  Google Scholar 

  15. Kulkarni RD, Schaefer MR, Golden SS: Transcriptional and posttranscriptional components of psbA response to high light intensity in Synechococcus sp. strain PCC 7942. J Bact 174: 3775–3781 (1992).

    PubMed  Google Scholar 

  16. Li R, Dickerson NS, Mueller UW, Golden SS: Specific binding of Synechococcus sp. PCC 7942 proteins to the enhancer element of psbAII required for high-light-induced expression. J Bact 177: 508–516 (1995).

    PubMed  Google Scholar 

  17. Li R, Golden SS: Enhancer activity of the light-responsive regulatory elements in the untranslated leader regions of cyanobacterial psbA genes. Proc Natl Acad Sci USA 90: 11678–11682 (1993).

    PubMed  Google Scholar 

  18. Nanba O, Satoh K: Isolation of a photosystem II reaction centre consisting of D-1 and D-2 proteins and cytochrome b-559. Proc Natl Acad Sci USA 84: 109–112 (1987).

    Google Scholar 

  19. Rao VSK, Brand JJ, Myers J: Cold shock syndrome in Anacystis nidulans. Plant Physiol 59: 965–969 (1977).

    Google Scholar 

  20. Richardson JP: Rho-dependent transcription termination. Biochim Biophys Acta 1048: 127–138 (1990).

    PubMed  Google Scholar 

  21. Richardson JP: Preventing the synthesis of unused transcripts by Rho factor. Cell 64: 1047–1049 (1991).

    PubMed  Google Scholar 

  22. Ruteshouser EC, Richardson JP: Identification and characterization of transcription termination sites in the Escherichia coli lacZ gene. J Mol Biol 208: 23–43 (1989).

    PubMed  Google Scholar 

  23. Schaefer MR, Golden SS: Differential expression of members of a cyanobacterial psbA gene family in response to light. J Bact 171: 3973–3981 (1989a).

    PubMed  Google Scholar 

  24. Schaefer MR, Golden SS: Light availability influences the ratio of two forms of D1 in cyanobacterial thylakoids. J Biol Chem 264: 7412–7417 (1989b).

    PubMed  Google Scholar 

  25. Scherrer K, Darnell JE: Sedimentation characteristics of rapidly labelled RNA from HeLa cells. Biochem Biophys Res Commun 7: 486–490 (1962).

    PubMed  Google Scholar 

  26. Shimamoto T, Noguchi K, Kuroda M, Tsuda M and Tsuchiay T: Transcriptional attenuation and differential mRNA stability in the regulation of the Escherichia coli Melibiose operon. J Biochem 115: 1185–1189 (1994).

    PubMed  Google Scholar 

  27. Sippola K, Kanervo E, Murata N, Aro E-M: The genetically engineered increase in fatty acid unsaturation in Synechococcus sp. PCC 7942 allows D1-protein form exchange and sustenance of PSII activity at low temperature. Eur J Biochem 251: 641–648 (1998).

    PubMed  Google Scholar 

  28. Soitamo AJ, Zhou G, Clarke AK, Öquist G, Aro E-M, Gustafsson P: Over-production of the D1 protein of photosystem II reaction centre in the cyanobacterium Synechococcus sp. PCC 7942. Plant Mol Biol 26: 709–721 (1994).

    PubMed  Google Scholar 

  29. Soitamo AJ, Zhou G, Clarke AK, Öquist G, Gustafsson P, Aro E-M: Over-production of the D1:2 protein makes Synechococcus cells more tolerant to photoinhibition of photosystem II. Plant Mol Biol 30: 467–478 (1996).

    PubMed  Google Scholar 

  30. Stanssens P, Remaut E, Fiers W: Inefficient translation causes premature transcription termination. Cell 44: 711–718 (1986).

    PubMed  Google Scholar 

  31. Trebst A: The topology of the plastoquinone and herbicide binding peptides of photosystem II in the thylakoid membrane. Z Naturforsch 41c 240–245 (1986).

    Google Scholar 

  32. Tomioka N, Shinozaki K, Sugiura M: Molecular cloning and characterization of ribosomal RNA genes from a blue-green alga,Anacystis nidulans. Mol Gen Genet 184: 358–363 (1981).

    Google Scholar 

  33. Tsurushita N, Shigesada K and Imai M: Mutant rho factors with increased transcription termination activities. Functional correlations of the primary and secondary polynucleotide binding sites with the efficiency and site-selectivity of rhodependent termination. J Mol Biol 210: 23–37 (1989).

    PubMed  Google Scholar 

  34. Tyystjärvi T, Aro E-M, Jansson C, Mäenpää P: Changes in amino acid sequence in PEST-like area and QEEET-motif affect degradation rate of D1 polypeptide in photosystem II. Plant Mol Biol 25: 517–526 (1994).

    PubMed  Google Scholar 

  35. Wek RC, Sameshima JH and Hatfield GW: Rho-dependent transcriptional polarity in the ilvGMEDA operon of wild-type Escherichia coli K12. J Biol Chem 262: 15256–15261 (1987).

    PubMed  Google Scholar 

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Soitamo, A., Sippola, K. & Aro, EM. Expression of psbA genes produces prominent 5′ psbA mRNA fragments in Synechococcus sp. PCC 7942. Plant Mol Biol 37, 1023–1033 (1998). https://doi.org/10.1023/A:1006077824075

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  • DOI: https://doi.org/10.1023/A:1006077824075

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