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Microbiology 146 (2000), 2877-2889
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Microbiology (2000), 146, 2877-2889.
© 2000 Society for General Microbiology

Genetics and Molecular Biology

Stress responses of Synechocystis sp. strain PCC 6803 mutants impaired in genes encoding putative alternative sigma factors

Jana Huckauf1, Chris Nomura2, Karl Forchhammer3 and Martin Hagemann1

Universität Rostock, FB Biologie, Institut für Molekulare Physiologie und Biotechnologie, Doberaner Str. 143,D-18051 Rostock, Germany1
Department of Biochemistry and Molecular Biology, The Pennsylvania State University, University Park, PA 16802, USA2
Justus-Liebig-Universität Giessen, Institut für Mikrobiologie und Molekularbiologie, Frankfurter Str. 107,D-35392 Giessen, Germany3

Author for correspondence: Martin Hagemann. Tel: +49 381 4942076. Fax: +49 381 4942079. e-mail: mh{at}bio4.uni-rostock.de

In the complete genome sequence of the cyanobacterium Synechocystis sp. strain PCC 6803 [Kaneko et al. (1996 ). DNA Res 3, 109–136] genes were identified encoding putative group 3 {sigma}-factors SigH (Sll-0856), SigG (Slr-1545) and SigF (Slr-1564) and the regulatory protein RsbU (Slr-2031). Mutations in these genes were generated by interposon mutagenesis to study their importance in stress acclimation. For the genes sigH, sigF and rsbU, the loci segregated completely. However, attempts to mutagenize the sigG locus resulted in merodiploids. Under standard growth conditions only minor differences were detected between the mutants and wild-type. However, cells of the RsbU mutant showed a clear defect in regenerating growth after a nitrogen- and sulphur-starvation-induced stationary phase. After applying salt, heat and high-light shocks, stress protein synthesis was analysed by means of one- and two-dimensional electrophoresis. Cells of the SigF mutant showed a severe defect in the induction of salt stress proteins. Although the acclimation to moderate salt stress up to 684 mM NaCl was not significantly changed in this mutant, its ability to acclimate to higher concentrations of NaCl was reduced. Northern blot experiments showed a constitutive expression of the rsbU and sigF genes. The expression of the sigH gene was found to be stress-stimulated, particularly in heat-shocked cells, whilst that of sigG was transiently decreased under stress conditions. Possible functions of these regulatory proteins in stress acclimation of Synechocystis cells are discussed.

Keywords: cyanobacteria, environmental stress, sigma factors, stationary phase, stress proteins

Abbreviations: 1D, one-dimensional; 2D, two-dimensional; Km, kanamycin; PSII, photosystem II; WT, wild-type




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