Search

Acid stress is an environmental problem for plants and fresh water cyanobacteria like the filamentous, heterocyst forming species Anabaena sp. PCC 7120 (hereafter Anabaena sp.). Heterocyst differentiation, cell–cell communication and nitrogen fixation has been deeply studied in this model organism, but little is known about the cellular response of Anabaena sp. to decreased pH values, causing acid stress. ATP-binding cassette (ABC) transporters are involved in acid stress response in other bacteria, by exporting proteins responsible for survival under acidification. The genome of Anabaena sp. encodes numerous ABC transporter components, whose function is not known yet. Here, we describe the function of the gene all5304 encoding a protein with homology to membrane fusion proteins of tripartite efflux pumps driven by ABC transporters like HlyBD-TolC of Escherichia coli. The all5304 mutant shows less resistance against low pH, even though the expression of the gene is independent from the pH of the medium. We compared the exoproteome of the wild type and mutant cultures and identified three proteins-candidate substrates of the putative transporter. Including the in silico analysis of All5304, our results suggest that All5304 functions as part of an efflux pump, secreting of a protein necessary for acid tolerance in Anabaena sp. [ABSTRACT FROM AUTHOR]

Plastocyanin, encoded by the petE gene, can transfer electrons to photosystem I ( PSI) and cytochrome c oxidase during photosynthetic and respiratory metabolism in cyanobacteria. We constructed a petE mutant of Synechocystis sp. strain PCC 6803 and investigated its phenotypic properties under different light conditions. When cultured under continuous light, inactivation of petE accelerated the plastoquinone pool reoxidation, slowed the reoxidation rate of the primary quinone-type acceptor, and decreased the connectivity factor between the individual photosystem II ( PSII) photosynthetic units. Compared with the wild-type control, the petE mutant showed a decrease in its PSI/ PSII fluorescence ratio and an increase in its dark respiration rate. When cultured under a light-dark photoperiod, the petE mutation caused an increase in the phycocyanin to chlorophyll ratio. Consequently, the mutant line was a darker blue than its wild-type counterpart. Moreover, the petE mutation increased the efficiency of light capture, nonphotochemical quenching, and linear electron transport activity, but decreased the functional absorption cross section of PSII. These results suggest that plastocyanin is involved in regulating the redox state of the photosynthetic electron transfer chain, and the petE mutation can induce interesting phenotypic properties that are specific to the light-dark photoperiod. [ABSTRACT FROM AUTHOR]

It has been shown that efficient functioning of photosynthesis and respiration in the cyanobacterium Synechocystis PCC 6803 requires the presence of either cytochrome c6 or plastocyanin. In order to check whether the blue copper protein plastocyanin can act as electron donor to cytochrome c oxidase, we investigated the intermolecular electron transfer kinetics between plastocyanin and the soluble CuA domain (i.e. the donor binding and electron entry site) of subunit II of the aa3-type cytochrome c oxidase from Synechocystis. Both copper proteins were expressed heterologously in Escherichia coli. The forward and the reverse electron transfer reactions were studied yielding apparent bimolecular rate constants of (5.1±0.2)×104 M−1s−1 and (8.5±0.4)×105 M−1s−1, respectively (20 mM phosphate buffer, pH 7). This corresponds to an apparent equilibrium constant of 0.06 in the physiological direction (reduction of CuA), which is similar to Keq values calculated for the reaction between c-type cytochromes and the soluble fragments of other CuA domains. The potential physiological role of plastocyanin in cyanobacterial respiration is discussed. [Copyright &y& Elsevier]

The effects of various NaCl and sorbitol concentrations in the growth medium on polyamine content and on two enzymes of the polyamine biosynthesis pathway, arginine decarboxylase (ADC) and S-adenosyl methionine decarboxylase (SAMDC), were investigated in the unicellular cyanobacterium Synechocystis sp. PCC 6803. Synechocystis cells showed no difference in growth rate when the concentration of NaCl was raised up to 550 mM. The growth rate decreased at 300 mM sorbitol, and complete inhibition of growth occurred at concentrations of ≥700 mM sorbitol. Salt stress induced a moderate increase in the total cellular polyamine content, spermine in particular. Osmotic stress caused an apparent increase in the total cellular polyamine content with a marked increase of spermidine induced by 700 mM sorbitol. Importantly, a low level of spermine, which so far has never been detected in cyanobacteria, could be found in Synechocystis sp. PCC 6803. ADC, a key enzyme for putrescine synthesis, was unaffected by salt stress but showed a six-fold increase in enzyme activity upon osmotic stress imposed by 700 mM sorbitol. SAMDC, another important enzyme for spermidine and spermine synthesis, responded to salt and osmotic stresses similarly to the pattern observed for ADC. An analysis by reverse transcription-polymerase chain reaction revealed an increase of ADC mRNA level in cells under salt and osmotic stresses. Most importantly, the increase of ADC mRNA was attributed to its slower turnover rate under both stress conditions. Interestingly, the samdc gene(s) of Synechocystis appear to be unique since comparisons with known gene sequences from other organisms resulted in no homologous sequences identified in the Synechocystis genome. [Copyright &y& Elsevier]

Biochemical responses to cold and osmotic stresses overlap because each decreases the availability of free water. Since RNA-binding proteins are known to accumulate following cold stress and play key roles in regulating transcription termination, the effect of osmotic stress on expression of RNA-binding proteins was examined. The transcript levels of four genes encoding RNA-binding proteins (rbpA, rbpB, rbpC and rbpD) were monitored in Anabaena sp. PCC 7120 cultures supplemented with ammonium ions or growing under nitrogen-fixing conditions. Steady-state transcript levels of all four genes increased transiently in response to a temperature shift from 30 to 20°C under both nitrogen regimes. Osmotic stress also enhanced rbpB, rbpC and rbpD gene expression in ammonium grown cultures. In the absence of a combined nitrogen source, osmotic stress repressed the short-term induction of rbp gene expression. The accumulation of RNA-binding proteins did not follow transcript levels, but remained high 24 h after stress initiation. It is concluded that nitrogen nutrition modulates the stress-responsive regulation of RNA-binding proteins in cyanobacteria, providing a potential mechanism to integrate environmental and developmental signals. [Copyright &y& Elsevier]

Two-component signal transduction systems, composed of histidine sensory kinases and response regulators, constitute a key element of the mechanism by which bacteria sense and acclimatize to changes in their environment. The availability of whole genome sequences permits a detailed analysis of these genes in cyanobacteria. In the present paper, we focus mainly on Prochlorococcus MED4, a strain adapted to surface oceanic conditions, for which six putative response regulators (rer) and five putative histidine kinases (hik) were identified. These numbers are comparable to those found in the other marine picocyanobacteria but much lower than those found in freshwater cyanobacteria. Moreover, the diversity of these genes is low in Prochlorococcus since most histidine kinases are related to a single group (type I) and most response regulators to a single family (OmpR). Under standard conditions, quantitative reverse transcription polymerase chain reaction revealed that one hik (hik03) and two rer (rer04 and rer05) genes were expressed at relatively high levels compared to the other two-component system genes. In response to high light exposure, a moderate increase (>5-fold) was observed in the expression of some putative rer genes (rer01, rer04, rer05, and rer06), whereas a smaller increase (<3-fold) in hik03 and hik04 mRNA levels was detected. In contrast, both cold and heat shocks decreased rather than increased the expression of most hik and rer genes. [Copyright &y& Elsevier]

When growing on N2 as sole nitrogen source, the filamentous cyanobacterium Anabaena sp. PCC 7120 forms N2 fixing heterocysts in a semi-regular pattern. To identify genes involved in heterocyst differentiation we characterised five transposon-generated mutants that were not able to form mature heterocysts. After recovering the transposon together with the flanking region of the Anabaena chromosome the affected genes were identified. Four of the genes could be involved in formation of the heterocyst-specific envelope: alr2887, encoding a probable outer membrane efflux protein, alr3698, a glycosyl transferase, all4388, a putative periplasmic polysaccharide export protein and alr5357, the formerly described gene hglB/hetM, encoding a fatty-acid synthetase. Another gene, all0049/mutS2, may be important in one of the genome rearrangements that occur during heterocyst differentiation. By transcriptional fusion to reporter genes luxAB differential expression of alr2887, alr3698 and alr5357 could be monitored during heterocyst differentiation. [Copyright &y& Elsevier]

The leader peptide of the major secreted protein PilA1 of the cyanobacterium Synechocystis sp. strain PCC 6803 and several artificial leader peptides have been used to study secretion of the reporter protein lichenase to the culture medium. The strains of Synechocystis carrying lichenase with the leader sequences of PilA and with the leader sequence of Slr2016 efficiently secreted the reporter protein. The artificial leader sequence that was characterized by the overall positive charge (as PilA1 and Slr2016 leaders) also allowed secretion. The artificial leader with negative charge, however, did not allow secretion of the reporter protein. Moreover, no secreted proteins have been isolated from this strain using conventional techniques for preparation of secreted proteins. These data suggest that the general secretion pathway in cyanobacteria, at least for pilins, recognizes the overall charge of the leader sequences, and operates in a sequence-non-specific manner. [Copyright &y& Elsevier]

Wild-type strains of the cyanobacterium Planktothrix rubescens have a cluster of gas vesicle (gvp) genes with repeats of alternating gvpA and gvpC. The gvpC occurs in three length variants, all with the same 3′-sequence, ΩC. Spontaneous non-buoyant mutants had lost some of the alternating gvpAC copies and their gvpC genes had a novel 3′-end sequence, ΨC; additional gvpC genes terminating in this sequence were also found in the wild-type and representatives of other GV genotypes. Alleles of gvpC terminating in ΨC occurred only at the downstream ends of the gvpAC clusters investigated; all other gvpCs terminated in ΩC. Mutants of strains with the GV3 genotype produced only 30–50% of the gas vesicles present in the wild-type; their gas vesicles had lower mean critical pressures (0.70–0.78 MPa) than those in the wild-type (1.05–1.10 MPa). [Copyright &y& Elsevier]

Mycosporine-like amino acids (MAAs) were extracted from a unicellular cyanobacterium (Euhalothece sp.) isolated from a gypsum crust on the bottom of a hypersaline saltern pond in Eilat, Israel. When grown at high light intensities, this isolate contained high concentrations of two MAAs, one showing maximum optical density at 331 nm and one at 362 nm. The compound absorbing at 331 nm was purified by preparative high performance liquid chromatography, and its structure was elucidated by one-dimensional (1H and 13C) and two-dimensional nuclear magnetic resonance, mass spectrometry and amino acid analysis, and identified as mycosporine-2-glycine. This is the first report of mycosporine-2-glycine in cyanobacteria. [Copyright &y& Elsevier]