SigC sigma factor is involved in acclimation to low inorganic carbon at high temperature in Synechocystis sp. PCC 6803.

Inactivation of the sigC gene (sll0184), encoding the group 2 sigma factor SigC, leads to a heat-sensitive phenotype of Synechocystis sp. PCC 6803. Cells of the DeltasigC strain grew poorly at 43 degrees C at pH 7.5 under ambient CO(2) conditions. Addition of inorganic carbon in the form of 3 % CO(2) or use of an alkaline growth medium (pH 8.3) restored the growth of the DeltasigC strain at 43 degrees C. These treatments compensate for the low concentration of inorganic carbon at high temperature. However, addition of organic carbon as glucose, pyruvate, succinate or 2-oxoglutarate did not restore growth of the DeltasigC strain at 43 degrees C. In the control strain, the amount of the SigC factor diminished after prolonged incubation at 43 degrees C if the pH of the growth medium was 7.5 or 6.7. Under alkaline conditions, the amount of the SigC factor remained constant at 43 degrees C and cells of the control strain grew better than at pH 7.5 or pH 6.7. The pH dependence of high-temperature growth was associated with changes in photosynthetic activity, indicating that the SigC factor is involved in adjustment of photosynthesis according to the amount of available inorganic carbon. Our results indicate that acclimation to low inorganic carbon is a part of acclimation to prolonged high temperature and that the SigC factor has a central role in this acclimation.

Characterization of single and double inactivation strains reveals new physiological roles for group 2 sigma factors in the cyanobacterium Synechocystis sp. PCC 6803.

Cyanobacteria are eubacteria that perform oxygenic photosynthesis like plants. The initiation of transcription, mediated by the RNA polymerase holoenzyme, is the main determinant of gene regulation in eubacteria. The sigma factor of the RNA polymerase holoenzyme is responsible for the recognition of a promoter sequence. In the cyanobacterium Synechocystis sp. PCC 6803, the primary sigma factor, SigA, is essential for cell viability. The SigB, SigC, SigD, and SigE factors show significant sequence similarity with the SigA factor but are nonessential. In this study, we have used homology modeling to construct a three-dimensional model of Synechocystis RNA polymerase holoenzyme and all group 1 and 2 sigma factors. According to the models, the overall three-dimensional structures of group 1 and 2 sigma factors are similar, the SigB and SigD factors being the most similar ones. In addition, we have constructed a complete set of group 2 sigma factor double inactivation strains, DeltasigBC, DeltasigBD, DeltasigBE, DeltasigCD, DeltasigCE, and DeltasigDE. All double mutants grow well under standard conditions, but differences are observed in stress conditions. The transition from lag phase to exponential growth is slow in the DeltasigBD strain, and all strains lacking the SigD factor were found to be sensitive to bright light. Furthermore, all group 2 sigma factors were found to be involved in acclimation to salt- or sorbitol-induced osmotic stresses.

Sigma factor SigC is required for heat acclimation of the cyanobacterium Synechocystis sp. strain PCC 6803.

The role of the primary-like sigma factor SigC was studied in Synechocystis. Under high temperature stress (48 degrees C) the DeltasigC inactivation strain showed a lower survival rate than the control strain. The DeltasigC strain grew poorly at 43 degrees C in liquid cultures under normal air. However, change to 3% CO(2) enhanced growth of DeltasigC at 43 degrees C. Differences in expression of many genes related to the carbon concentrating mechanisms between the control and the DeltasigC strain were recorded with a genome-wide DNA microarray. We suggest that low solubility of CO2 at high temperature is one of the factors contributing to the poor thermotolerance of the DeltasigC strain.

The SigB sigma factor mediates high-temperature responses in the cyanobacterium Synechocystis sp. PCC6803.

The sigma factors of RNA polymerase play central roles when bacteria adapt to different environmental conditions. We studied heat-shock responses in the cyanobacterium Synechocystis sp. PCC6803 using the sigma factor inactivation strains deltasigB, deltasigD and deltasigBD. The SigB factor was found to be important for short-term heat-shock responses and acquired thermotolerance. The normal high-temperature induction of the hspA gene depended on the SigB factor. The SigD sigma factor had a role in high-temperature responses as well, and the double inactivation strain deltasigBD grew more slowly at 43 degrees C than the deltasigB and deltasigD strains.

Evidence for the role of the oxygen-evolving manganese complex in photoinhibition of Photosystem II.

Photoinhibition of PSII occurs at the same quantum efficiency from very low to very high light, which raises a question about how important is the rate of photosynthetic electron transfer in photoinhibition. We modulated electron transfer rate and light intensity independently of each other in lincomycin-treated pea leaves and in isolated thylakoids, in order to elucidate the specific effects of light and PSII electron transport on photoinhibition. Major changes in the rate of electron transport caused only small changes in the rate of photoinhibition, suggesting the existence of a significant photoinhibitory pathway that contains an electron-transfer-independent phase. We compared the action spectrum of photoinhibition with absorption spectra of PSII components that could function as photoreceptors of the electron-transfer-independent phase of photoinhibition and found that the absorption spectra of Mn(III) and Mn(IV) compounds resemble the action spectrum of photoinhibition, showing a steep decrease from UV-C to blue light and a low visible-light tail. Our results show that the release of a Mn ion to the thylakoid lumen is the earliest detectable step of both UV- and visible-light-induced photoinhibition. After Mn release from the oxygen-evolving complex, oxidative damage to the PSII reaction center occurs because the Mn-depleted oxygen-evolving complex cannot reduce P680+ normally.

Expression of primary sigma factor (PSF) and PSF-like sigma factors in the cyanobacterium Synechocystis sp. strain PCC 6803.

Large amounts of sigA mRNA, encoding the primary sigma factor (PSF) in Synechocystis sp. strain PCC 6803, accumulated under standard growth conditions, while stress conditions like heat or high salinity led to a rapid decrease in sigA mRNA content. The sigB, sigC, sigD, and sigE genes, encoding PSF-like sigma factors, were under strict physiological control.

Action spectrum of psbA gene transcription is similar to that of photoinhibition in Synechocystis sp. PCC 6803.

The photosystem II (PSII) reaction center protein D1 undergoes rapid light-dependent turnover, which is caused by photoinhibition. To identify the photoreceptor(s) involved in the light-dependent expression of the psbA gene encoding the D1 protein, we determined the action spectra of psbA transcription, PSII activity, photosynthesis and photoinhibition in Synechocystis sp. PCC 6803. In accordance with its phycobilisome antenna, PSII showed the highest activity in the spectral region from yellow to red and only low activity in the ultraviolet-A (UV-A) to green region. Photoinhibition, in turn, was fastest in UV-A to violet light and a minor peak was found in the orange region. The action spectrum of psbA transcription resembled closely that of photoinhibition, suggesting that photoinhibition creates a signal for up-regulation of the psbA gene.