[Inversion of phototaxis in cells of Synechocystis sp. PCC 6803 determined by a mutation in the regulatory gene prqR].

Phototaxis, positive (movement toward the light source) or negative (from the light source) mediates the adaptation of cyanobacteria to varying wave lengths and illumination intensity. The transcription regulator PrqR of the family TetR is known as a repressor of the prqRA operon controlling resistance to the oxidative stress inducer methyl viologen in the cyanobacterium Synechocystis sp. PCC6803. However, it was shown in this work that mutation prqRL17Q affecting the DNA-binding domain of the PrqR protein, which causes derepression of the prqRA operon and enhances cell resistance to methyl viologen, additionally determines negative phototaxis induced with daylight and red light of low intensity. The inactivation of gene prqA did not affect cell motility in mutant PqR carrying mutation prqRL17Q and in the wild-type strain characterized by positive phototaxis appearing in response to the light of low intensity. Moreover, a mutant with deletion prqR did not differ from the wild-type strain with respect to phototaxis type suggesting that the specificity of the regulator protein was changed in cells carrying prqRL17Q mutation. Note that changes in transcription of pilA genes that control biogenesis of pili providing for cell motility were not detected in mutant PqR, and, in agreement with data of atomic force microscopy, the type of pili formation is identical in prqR mutants and the wild-type strain. Meanwhile, mutant PqR manifested a decrease in transcription of gene taxD1 encoding the photoreceptor of red light that is required for the positive phototaxis of cyanobacteria. These data imply that mutation prqRL17Q changes the specificity of the PrqR repressor protein and thereby affects the regulation of phototaxis at the level of photoperception and signal transduction in cells.

[The sll0886 gene, controlling light-activated heterotrophic growth, is involved in regulating phototaxis in cyanobacterium Synechocystis sp. PCC 6893].

The sll0886 gene, controlling light-activated heterotrophic growth (LAHG), was tested for the role in regulating phototaxis in cyanobacterium Synechocystis sp. PCC 6803. Insertional inactivation of the gene in the genome of a wildtype strain did not affect positive (toward light) or negative (away from high light) phototaxis. However, cells lost motility when sll0886 inactivation was combined with the prqRL17Q mutation, which determined negative phototaxis at low light. Immotile cells with the prqRL17Q mutation and the inactivated sll0886 gene did not display any defect in the formation of type IV pili, essential for phototaxis. Hence, the function, rather than biogenesis, of pili was affected. It was concluded that the sll0886 gene, coding for a TPR family protein, is involved in controlling negative phototaxis of cyanobacteria at the level of photoreception and signal transduction and that its role is mediated by the unidentified redundant gene whose function is suppressed by the prqRL17Q mutation.

[Trans- and cis-acting autorepressors of the prqR gene in Synechocystis cyanobacteria sp. PCC6803]. / Trans- i tsis-deistvuiushchaia avtorepressiia gena prqR u tsianobakterii Synechocystis sp. PCC 6803.

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 inductor methyl viologen (MV) in cyanobacterium Synechocystis sp. PCC 6803. The wild-type (WT) prqR showed 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 C-terminal domain in transcriptional repression by PrqR. Missense mutation C134S altering 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 WT and mutant prqR alleles in Synechocystis cells revealed a cis-dominant character of the alteration of prqR autoregulation. Stimulation of in cis autorepression of prqR was assumed to contribute to the induction of systems protecting cyanobacteria from oxidative stress.