Abstract
Genetic analysis of the allele interactions was carried out with the use of recombinant plasmids and reporter genes to study the autorepressor function of prqR, which negatively regulates the prqR–prqA operon and the response to oxidative stress inducer methyl viologen (MV) in cyanobacterium Synechocystis sp. PCC 6803. The wild-type prqR cloned in Escherichia colishowed negative autoregulation and suppressed in trans the derepressed mutant alleles. Frameshift mutation C134fs, which was introduced in prqR by site-directed mutagenesis, impaired the autoregulation, implicating the PrqR C-terminal domain in transcriptional repression. Missense mutation C134S, changing the only redox-sensitive Cys of PrqR, had no effect on prqR expression, indicating that oxidation and consequent disulfide bridging of two PrqR molecules was not responsible for MV-induced autorepression of prqR. Analysis of the prqR–prqA deletion derivatives lacking the promoter and most of prqR revealed weak uncontrollable expression of reporter cat, testifying to the existence of a constitutive promoter in prqA responsible for MV resistance. The interaction of the wild-type and mutant prqR alleles in Synechocystis cells revealed a cis-dominant character of the impairment of prqR autoregulation. Stimulation of in cis autorepression of prqR was assumed to contribute to the induction of systems protecting cyanobacteria against oxidative stress.
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REFERENCES
Farr S.B., Kogoma T. 1991. Oxidative stress responses in Escherichia coli and Salmonella typhimurium. Microbiol. Rev. 55, 561–585.
Asada K. 1994. Production and action of active oxygen species in photosynthesis tissues. In: Causes of photooxidative stress and amelioration of defense systems in plants. Foyer C.H., Mullineaux P.M. Eds. Boca Raton, Fla.: CRC Press, Inc., 77–104.
Obinger C., Regelsberger G., Strasser G., Burner U., Peschek G.A. 1997. Purification and characterization of a homodimeric catalase-peroxidase from the cyanobacterium Anacystis nidulans. Biochem. Biophys. Res. Commun. 235, 545–552.
Thomas D.J., Avenson T.J., Thomas J.B., Herbert S.K. 1998. A cyanobacterium lacking iron superoxide dismutase is sensitized to oxidative stress induced with methyl viologen but is not sensitized to oxidative stress induced with norflurazon. Plant Physiol. 116, 1593–1602.
Tichy M., Vermaas W. 1999. In vivo role of catalase-per-oxidase in Synechocystis sp. strain PCC 6803. J. Bacteriol. 181, 1875–1882.
Gaber A., Tamoi M., Takeda T., Nakano Y., Shigeoka S. 2001. NADPH-dependent glutathione peroxi-dase-like proteins (Gpx-1, Gpx-2) reduce unsaturated fatty acid hydroperoxides in Synechocystis PCC 6803. FEBS Lett. 499, 32–36.
Storz G., Imlay J.A. 1999. Oxidative stress. Current Opin. Microbiol. 2, 188–194.
Pomposiello P., Demple B. 2001. Redox-operated genetic switches: the SoxR and SoxS transcription factors. Trends Biotechnol. 19, 109–114.
Sidoruk K., Melnik V., Babykin M., Cerff R., Shestakov S. 1999. Cloning and molecular analysis of the gene prqR controlling resistance to paraquat in Synechocystis sp. PCC 6803. In: The Phototrophic Prokaryotes. Peschek G.A., Loffelhardt W., Schmetterer G. Eds. N.Y.: Kluwer Acad. / Plenum Publ., 715–718.
Babykin M.M., Sidoruk K.V., Zinchenko V.V., Nefedova L.N., Cerff R., Shestakov S.V. 2003. On the involvement of the prqR regulatory gene in methyl viologen resistance of cyanobacterium Synechocystis sp. PCC 6803. Genetika. 39, 25–32.
Nefedova L.N., Fantin Yu.S., Zinchenko V.V., Babykin M.M. 2003. The prqA and mvrA genes coding for transporter proteins control the methyl viologen sensitivity of cyanobacterium Synechocystis sp. PCC 6803. Genetika. 39, 336–340.
Brown T.A. 1991. Molecular biology LabFax. Hames B.D., Rickwood D. Eds. Oxford, UK: BIOS Scientific Publishers Limited.
Sambrook J., Fritsch E.F., Maniatis T. 1989. Molecular Cloning: A Laboratory Manual, 2nd ed. Cold Spring Harbor, N.Y.: Cold Spring Harbor Laboratory Press.
Rippka R., Deruelles D.E., Waterbury J.B., Herdman M., Stanier R.Y. 1979. Generic assignments, strain histories and properties of pure cultures of cyanobacteria. J. Gen. Microbiol. 111, 1–61.
Grigorieva G., Shestakov S.V. 1982. Transformation in the cyanobacterium Synechocystis sp. 6803. FEMS Microbiol. Lett. 13, 367–370.
Zinchenko V.V., Babykin M.M., Shestakov S.V. 1984. Mobilization of non-conjugative plasmids into Rhodopseudomonas sphaeroides. J. Gen. Microbiol. 130, 1587–1590.
Holmes D.S., Quigley M. 1981. A rapid boiling method for the preparation of bacterial plasmids. Anal. Biochem. 114, 193–197.
Cyanobase Website. http://www.kazusa.or.jp/cyano/ Synechocystis.
Scholz P., Haring V., Wittmann-Liebold B., Ashman K., Bagdasarian M., Scherzinger E. 1989. Complete nucleotide sequence and gene organization of the broad-host-range plasmid RSF1010. Gene. 75, 271–288.
Schweizer H.P. 1993. Small broad-host-range gentamycin resistance gene cassettes for site-specific insertion and deletion mutagenesis. BioTechniques. 15, 831–833.
Hillen W., Berens C. 1994. Mechanisms underlying expression of Tn10 encoded tetracycline resistance. Annu. Rev. Microbiol. 48, 345–369.
DeLong A., Syvanen M. 1991. Transacting transposase mutant from Tn5. Proc. Natl. Acad. Sci. USA. 88, 6072–6076.
Masai H., Kaziro Y., Arai K. 1983. Definition of oriR, the minimum DNA segment essential for initiation of R1 plasmid replication in vitro. Proc. Natl. Acad. Sci. USA. 80, 6814–6818.
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Kirik, I.A., Zinchenko, V.V., Shestakov, S.V. et al. In Cis and In Trans Autorepression of the prqR Gene in Cyanobacterium Synechocystis sp. PCC 6803. Molecular Biology 37, 880–887 (2003). https://doi.org/10.1023/B:MBIL.0000008357.98784.63
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DOI: https://doi.org/10.1023/B:MBIL.0000008357.98784.63