The gene sll0273 of the cyanobacterium Synechocystis sp. strain PCC6803 encodes a protein essential for growth at low Na+/K+ ratios

Mikkat, Stefan; Milkowski, Carsten; Hagemann, Martin

doi:10.1046/j.1365-3040.2000.00565.x

Cited by 26 publications

(22 citation statements)

References 44 publications

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“…Similar studies with Synechocystis sp. strain PCC 6803 did not detect any changes in the expression of Na ϩ /H ϩ antiporter genes under high-salt and alkaline conditions (12,15,22,23,26,33,35). However, a microarray analysis in which Synechocystis sp.…”

Section: Discussionmentioning

confidence: 98%

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Two Members of a Network of Putative Na ⁺ /H ⁺ Antiporters Are Involved in Salt and pH Tolerance of the Freshwater Cyanobacterium Synechococcus elongatus

2008

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Synechococcus elongatus strain PCC 7942 is an alkaliphilic cyanobacterium that tolerates a relatively high salt concentration as a freshwater microorganism. Its genome sequence revealed seven genes, nha1 to nha7 (syn_pcc79420811, syn_pcc79421264, syn_pcc7942359, syn_pcc79420546, syn_pcc79420307, syn_pcc79422394, and syn_pcc79422186), and the deduced amino acid sequences encoded by these genes are similar to those of Na ؉ /H ؉ antiporters. The present work focused on molecular and functional characterization of these nha genes encoding Na ؉ /H ؉ antiporters. Our results show that of the nha genes expressed in Escherichia coli, only nha3 complemented the deficient Na ؉ /H ؉ antiporter activity of the Na ؉ -sensitive TO114 recipient strain. Moreover, two of the cyanobacterial strains with separate disruptions in the nha genes (⌬nha1, ⌬nha2, ⌬nha3, ⌬nha4, ⌬nha5, and ⌬nha7) had a phenotype different from that of the wild type. In particular, ⌬nhA3 cells showed a high-salt-and alkaline-pH-sensitive phenotype, while ⌬nha2 cells showed low salt and alkaline pH sensitivity. Finally, the transcriptional profile of the nha1 to nha7 genes, monitored using the real-time PCR technique, revealed that the nha6 gene is upregulated and the nha1 gene is downregulated under certain environmental conditions.

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Section: Discussionmentioning

confidence: 98%

“…A low-Na ϩ requirement and high-pH lethality were also reported for a ⌬nhaS2 (orthologue of nha2) mutant of Synechocystis sp. strain PCC 6803 (33,65), reflecting conserved characteristics of these orthologues beyond amino acid sequence similarities.…”

Section: Discussionmentioning

confidence: 99%

Two Members of a Network of Putative Na ⁺ /H ⁺ Antiporters Are Involved in Salt and pH Tolerance of the Freshwater Cyanobacterium Synechococcus elongatus

2008

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“…Glutaredoxins are small ubiquitous glutathione-disulfide oxidoreductases that reduce disulfide bonds of target proteins and maintain the redox homoeostasis of Synechocystis cells, and disruption of a glutaredoxin coding gene ssr2061 in Synechocystis has been reported with reduced cell viability against oxidative stress (47). The sll0273 gene encoding a putative Na ϩ /H ϩ antiporter was found essential for growth at low Na ϩ /K ϩ ratios (48), and its mutant showed low tolerance to elevated pH in the BG11 medium (49). In our previous study, we found that the slr1829 gene encoding a putative poly(3-hydroxyalkanoate) synthase was inducted by butanol, and the enhanced production of carbon storage compound polyhydroxyalkanoates (PHAs) was one important mechanism for Synechocystis to combat against butanol stress (50).…”

Section: Fig 3 Reproducibility Between Biological Replicatesmentioning

confidence: 99%

A Transcriptional Regulator Sll0794 Regulates Tolerance to Biofuel Ethanol in Photosynthetic Synechocystis sp. PCC 6803

Song

Chen

Wang

et al. 2014

Molecular & Cellular Proteomics

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To improve ethanol production directly from CO 2 in photosynthetic cyanobacterial systems, one key issue that needs to be addressed is the low ethanol tolerance of cyanobacterial cells. Our previous proteomic and transcriptomic analyses found that several regulatory proteins were up-regulated by exogenous ethanol in Synechocystis sp. PCC6803. In this study, through tolerance analysis of the gene disruption mutants of the up-regulated regulatory genes, we uncovered that one transcriptional regulator, Sll0794, was related directly to ethanol tolerance in Synechocystis. Using a quantitative iTRAQ-LC-MS/MS proteomics approach coupled with quantitative real-time reverse transcription-PCR (RTqPCR), we further determined the possible regulatory network of Sll0794. The proteomic analysis showed that in the ⌬sll0794 mutant grown under ethanol stress a total of 54 and 87 unique proteins were down-and upregulated, respectively. In addition, electrophoretic mobility shift assays demonstrated that the Sll0794 transcriptional regulator was able to bind directly to the upstream regions of sll1514, slr1512, and slr1838, which encode a 16.6 kDa small heat shock protein, a putative sodium-dependent bicarbonate transporter and a carbon dioxide concentrating mechanism protein CcmK, respectively. The study provided a proteomic description of the putative ethanol-tolerance network regulated by the sll0794 gene, and revealed new insights on the ethanol-tolerance regulatory mechanism in Synechocystis. As the first regulatory protein discovered related to ethanol tolerance, the gene may serve as a valuable target for transcription machinery engineering to further improve ethanol tolerance in Synechocystis. All MS data have been deposited in the ProteomeXchange with identifier PXD001266

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“…The potassium ion content was estimated using flame photometry as described in great detail by Mikkat et al (2000). Inorganic ions were extracted by boiling the filters with 5 ml of double-distilled water for 10 min.…”

Section: + Determinationsmentioning

confidence: 99%

“…The residues were dissolved in defined volumes of water that contained 50 µg of sorbitol as an internal standard. Low-molecular mass compounds were quantitatively analyzed by gas chromatography (GC; Hagemann et al 2008) or high-performance liquid chromatography (HPLC; Hagemann et al 2005).The potassium ion content was estimated using flame photometry as described in great detail by Mikkat et al (2000). Inorganic ions were extracted by boiling the filters with 5 ml of double-distilled water for 10 min.…”

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confidence: 99%

Salt acclimation of Nodularia spumigena CCY9414—a cyanobacterium adapted to brackish water

Möke¹,

Wasmund²,

Bauwe³

et al. 2013

Aquat. Microb. Ecol.

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The toxic cyanobacterium Nodularia spumigena regularly forms large surface blooms in the central Baltic Sea. The Baltic Sea is characterized by a salinity gradient. We analyzed the salt acclimation of the strain N. spumigena CCY9414, the only Nodularia strain with a known genome sequence. N. spumigena CCY9414 showed a rather low salt tolerance range, displaying a growth optimum at 12.5 g NaCl l −1. Sucrose was identified as the major compatible solute. The expression of the sucrose-phosphate synthase gene was salt-stimulated, which indicates that the salt-induced sucrose accumulation could be regulated at the transcriptional level. Potassium ions and glutamate were also accumulated in Nodularia cells, especially at high salinities when sucrose levels were rather low. Our results indicate that N. spumigena CCY9414 represents a truly brackish-water-adapted cyanobacterial strain. KEY WORDS: Compatible solute · Glutamate · Potassium · SucroseResale or republication not permitted without written consent of the publisher Aquat Microb Ecol 70: 207-214, 2013 west connection to the North Sea and almost freshwater conditions in the Northern and Eastern edges (Samuelsson 1996). The distribution of Nodularia spumigena could depend on the external salinity because blooms occur mostly in the central Baltic Sea characterized by brackish waters around 8 practical salinity units (PSU) (Feistel et al. 2010). To adjust the internal osmotic potential to changing salinities, cyanobacteria are known to accumulate specific classes of compatible solutes (Hagemann 2011). Members of the group Nostocales, such as the model strain Anabaena sp. PCC 7119, usually show optimal growth under fresh water conditions and can acclimate to brackish water salinities by the accumulation of sucrose, which is catalyzed by sucrose-phosphate synthase (SPS) (Porchia & Salerno 1996).Compared to freshwater Anabaena strains, Nodularia spumigena seems to prefer brackish water conditions. To get deeper insight into the basic saltacclimation strategy of this ecologically important cyanobacterial genus, we aimed to investigate the salt response of N. spumigena under defined laboratory conditions. For this purpose, we used the Baltic Sea isolate N. spumigena CCY9414 (hereafter Nodularia CCY9414), taking advantage of its known genome sequence (GenBank accession no. AAVW 00000000; Voß et al. 2013). Genome searches revealed that Nodularia CCY9414 is potentially able to synthesize sucrose as well as trehalose as compatible solutes. Our results suggest that Nodularia CCY9414 is a brackish-water-adapted cyanobacterium because it grew optimally at salinities around 10 g NaCl l −1 and accumulated sucrose as the main compatible solute under these conditions. MATERIALS AND METHODS Strain and cultivationNodularia spumigena CCY9414 was obtained from the Culture Collection Yerseke (CCY) at the Netherlands Institute of Sea Research (NIOZ). This Baltic Sea strain was isolated from surface waters of ~9 PSU east of the island of Bornholm. It was taken into the strain ...

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The gene sll0273 of the cyanobacterium Synechocystis sp. strain PCC6803 encodes a protein essential for growth at low Na⁺/K⁺ ratios

Abstract: + /K+ ratios. This system may be part of a sodium cycle and may permit re-entry of Na + into the cells, if nutrient/Na + symporters are not functional or operating.

Cited by 26 publications

References 44 publications

Two Members of a Network of Putative Na ⁺ /H ⁺ Antiporters Are Involved in Salt and pH Tolerance of the Freshwater Cyanobacterium Synechococcus elongatus

Two Members of a Network of Putative Na ⁺ /H ⁺ Antiporters Are Involved in Salt and pH Tolerance of the Freshwater Cyanobacterium Synechococcus elongatus

A Transcriptional Regulator Sll0794 Regulates Tolerance to Biofuel Ethanol in Photosynthetic Synechocystis sp. PCC 6803

Salt acclimation of Nodularia spumigena CCY9414—a cyanobacterium adapted to brackish water

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The gene sll0273 of the cyanobacterium Synechocystis sp. strain PCC6803 encodes a protein essential for growth at low Na+/K+ ratios

Abstract: + /K+ ratios. This system may be part of a sodium cycle and may permit re-entry of Na + into the cells, if nutrient/Na + symporters are not functional or operating.

Cited by 26 publications

References 44 publications

Two Members of a Network of Putative Na + /H + Antiporters Are Involved in Salt and pH Tolerance of the Freshwater Cyanobacterium Synechococcus elongatus

Two Members of a Network of Putative Na + /H + Antiporters Are Involved in Salt and pH Tolerance of the Freshwater Cyanobacterium Synechococcus elongatus

A Transcriptional Regulator Sll0794 Regulates Tolerance to Biofuel Ethanol in Photosynthetic Synechocystis sp. PCC 6803

Salt acclimation of Nodularia spumigena CCY9414—a cyanobacterium adapted to brackish water

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The gene sll0273 of the cyanobacterium Synechocystis sp. strain PCC6803 encodes a protein essential for growth at low Na⁺/K⁺ ratios

Two Members of a Network of Putative Na ⁺ /H ⁺ Antiporters Are Involved in Salt and pH Tolerance of the Freshwater Cyanobacterium Synechococcus elongatus

Two Members of a Network of Putative Na ⁺ /H ⁺ Antiporters Are Involved in Salt and pH Tolerance of the Freshwater Cyanobacterium Synechococcus elongatus