Hypothetical chloroplast reading frame 51 encodes a photosystem I assembly factor in cyanobacteria.

Hypothetical chloroplast open reading frames (ycfs) are putative genes in the plastid genomes of photosynthetic eukaryotes. Many ycfs are also conserved in the genomes of cyanobacteria, the presumptive ancestors of present-day chloroplasts. The functions of many ycfs are still unknown. Here, we generated knock-out mutants for ycf51 (sll1702) in the cyanobacterium Synechocystis sp. PCC 6803. The mutants showed reduced photoautotrophic growth due to impaired electron transport between photosystem II (PSII) and PSI. This phenotype results from greatly reduced PSI content in the ycf51 mutant. The ycf51 disruption had little effect on the transcription of genes encoding photosynthetic complex components and the stabilization of the PSI complex. In vitro and in vivo analyses demonstrated that Ycf51 cooperates with PSI assembly factor Ycf3 to mediate PSI assembly. Furthermore, Ycf51 interacts with the PSI subunit PsaC. Together with its specific localization in the thylakoid membrane and the stromal exposure of its hydrophilic region, our data suggest that Ycf51 is involved in PSI complex assembly. Ycf51 is conserved in all sequenced cyanobacteria, including the earliest branching cyanobacteria of the Gloeobacter genus, and is also present in the plastid genomes of glaucophytes. However, Ycf51 has been lost from other photosynthetic eukaryotic lineages. Thus, Ycf51 is a PSI assembly factor that has been functionally replaced during the evolution of oxygenic photosynthetic eukaryotes.

Dehydration-Induced DnaK2 Chaperone Is Involved in PSII Repair of a Desiccation-Tolerant Cyanobacterium.

Maintaining the structural integrity of the photosynthetic apparatus during dehydration is critical for effective recovery of photosynthetic activity upon rehydration in a variety of desiccation-tolerant plants, but the underlying molecular mechanism is largely unclear. The subaerial cyanobacterium Nostoc flagelliforme can survive extreme dehydration conditions and quickly recovers its photosynthetic activity upon rehydration. In this study, we found that the expression of the molecular chaperone NfDnaK2 was substantially induced by dehydration, and NfDnaK2 proteins were primarily localized in the thylakoid membrane. NfDnaJ9 was identified to be the cochaperone partner of NfDnaK2, and their encoding genes shared similar transcriptional responses to dehydration. NfDnaJ9 interacted with the NfFtsH2 protease involved in the degradation of damaged D1 protein. Heterologous expression of NfdnaK2 enhanced PSII repair and drought tolerance in transgenic Nostoc sp. PCC 7120. Furthermore, the nitrate reduction (NarL)/nitrogen fixation (FixJ) family transcription factors response regulator (NfRre1) and photosynthetic electron transport-dependent regulator (NfPedR) were identified as putative positive regulators capable of binding to the promoter region of NfdnaK2 and they may mediate dehydration-induced expression of NfdnaK2 in N. flagelliforme Our findings provide novel insights into the molecular mechanism of desiccation tolerance in some xerotolerant microorganisms, which could facilitate future synthetic approaches to the creation of extremophiles in microorganisms and plants.

Sycrp2 Is Essential for Twitching Motility in the Cyanobacterium Synechocystis sp. Strain PCC 6803.

Two cAMP receptor proteins (CRPs), Sycrp1 (encoded by sll1371) and Sycrp2 (encoded by sll1924), exist in the cyanobacterium Synechocystis sp. strain PCC 6803. Previous studies have demonstrated that Sycrp1 has binding affinity for cAMP and is involved in motility by regulating the formation of pili. However, the function of Sycrp2 remains unknown. Here, we report that sycrp2 disruption results in the loss of motility of Synechocystis sp. PCC 6803, and that the phenotype can be recovered by reintroducing the sycrp2 gene into the genome of sycrp2-disrupted mutants. Electron microscopy showed that the sycrp2-disrupted mutant lost the pilus apparatus on the cell surface, resulting in a lack of cell motility. Furthermore, the transcript level of the pilA9-pilA11 operon (essential for cell motility and regulated by the cAMP receptor protein Sycrp1) was markedly decreased in sycrp2-disrupted mutants compared with the wild-type strain. Moreover, yeast two-hybrid analysis and a pulldown assay demonstrated that Sycrp2 interacted with Sycrp1 to form a heterodimer and that Sycrp1 and Sycrp2 interacted with themselves to form homodimers. Gel mobility shift assays revealed that Sycrp1 specifically binds to the upstream region of pilA9 Together, these findings indicate that in Synechocystis sp. PCC 6803, Sycrp2 regulates the formation of pili and cell motility by interacting with Sycrp1.IMPORTANCE cAMP receptor proteins (CRPs) are widely distributed in cyanobacteria and play important roles in regulating gene expression. Although many cyanobacterial species have two cAMP receptor-like proteins, the functional links between them are unknown. Here, we found that Sycrp2 in the cyanobacterium Synechocystis sp. strain PCC 6803 is essential for twitching motility and that it interacts with Sycrp1, a known cAMP receptor protein involved with twitching motility. Our findings indicate that the two cAMP receptor-like proteins in cyanobacteria do not have functional redundancy but rather work together.

Characterization of the sulfur-formation (suf) genes in Synechocystis sp. PCC 6803 under photoautotrophic and heterotrophic growth conditions.

MAIN CONCLUSION: The sulfur-formation ( suf ) genes play important roles in both photosynthesis and respiration of cyanobacteria, but the organism prioritizes Fe-S clusters for respiration at the expense of photosynthesis. Iron-sulfur (Fe-S) clusters are important to all living organisms, but their assembly mechanism is poorly understood in photosynthetic organisms. Unlike non-photosynthetic bacteria that rely on the iron-sulfur cluster system, Synechocystis sp. PCC 6803 uses the Sulfur-Formation (SUF) system as its major Fe-S cluster assembly pathway. The co-expression of suf genes and the direct interactions among SUF subunits indicate that Fe-S assembly is a complex process in which no suf genes can be knocked out completely. In this study, we developed a condition-controlled SUF Knockdown mutant by inserting the petE promoter, which is regulated by Cu2+ concentration, in front of the suf operon. Limited amount of the SUF system resulted in decreased chlorophyll contents and photosystem activities, and a lower PSI/PSII ratio. Unexpectedly, increased cyclic electron transport and a decreased dark respiration rate were only observed under photoautotrophic growth conditions. No visible effects on the phenotype of SUF Knockdown mutant were observed under heterotrophic culture conditions. The phylogenetic distribution of the SUF system indicates that it has a co-evolutionary relationship with photosynthetic energy storing pathways.

The hypothetical protein Ycf46 is involved in regulation of CO2 utilization in the cyanobacterium Synechocystis sp. PCC 6803.

MAIN CONCLUSION: The Ycf46 mutant of Synechocystis showed growth inhibition under low dissolved CO 2 conditions, suggesting a role for the Ycf46 protein in the process of photosynthetic CO 2 uptake and utilization. Hypothetical chloroplast open reading frame Ycf46 proteins are highly conserved in all cyanobacterial lineages and most algal chloroplast genomes, but their exact function is still unknown. In the cyanobacterium Synechocystis sp. PCC 6803, the Ycf46 encoding gene slr0374 is part of an operon (with slr0373 and slr0376) and responds to many environmental stresses. Transcript levels of the slr0373, slr0374 and slr0376 genes were increased under a low concentration of dissolved inorganic carbon (Ci). Compared with the wild type, the mutant lacking slr0374 showed growth arrest under Ci-deficient conditions but not under iron-deficient or low-light conditions. In addition, the mutant grew more slowly than the wild type under pH 6.0 conditions in which CO2 was the dominant Ci source, indicating the mutant cells had weak CO2 uptake and/or utilization ability. Supplying a high concentration of CO2 (5 %, v/v) to the mutant restored its phenotype to the wild type level. The photosynthetic activity of the mutant was inhibited to a lesser extent by a carbonic anhydrase inhibitor than that of the wild type, which specifically blocked CO2 uptake. Inactivation of slr0374 decreased expression of the ecaB gene and reduced carbonic anhydrase activity. A subcellular localization assay indicated that the Ycf46 protein was soluble. By co-immunoprecipitation assay using Slr0374 as a bait-protein, potential interacting proteins in the size range of 30 kDa were identified. These results suggest that the Ycf46 protein plays a role in the regulation of photosynthesis in cyanobacteria, especially in CO2 uptake and utilization.

Band Engineering and Phonon Engineering Effectively Improve n-Type Mg3Sb2 Thermoelectric Material Properties.

Mg3Sb2-based thermoelectric materials can convert heat and electricity into each other, making them a promising class of environmentally friendly materials. Further improving the electrical performance while effectively reducing the thermal conductivity is a crucial issue. In this paper, under the guidance of the oneness principle calculation, we designed a thermoelectric Zintl phase based on Mg3.2Sb1.5Bi0.5 doped with Tb and Er. Calculation results show that using Tb and Er as cationic site dopants effectively improves the electrical properties and reduces the lattice thermal conductivity. Experimental results confirmed the effectiveness of codoping and effectively enhanced thermoelectric performance. The most immense ZT value obtained by the Mg3.185Tb0.01Er0.005Sb1.5Bi0.5 sample was 1.71. In addition, the average Young's modulus of the Mg3.185Tb0.01Er0.005Sb1.5Bi0.5 sample is 51.85 GPa, and the Vickers hardness is 0.99 GPa. Under the same test environment, the material was subjected to 12 cycles in the temperature range of 323-723 K, and the average power factor error range was 1.8% to 2.1%, which is of practical significance for its application in actual device scenarios.

New insights into iron acquisition by cyanobacteria: an essential role for ExbB-ExbD complex in inorganic iron uptake.

Cyanobacteria are globally important primary producers that have an exceptionally large iron requirement for photosynthesis. In many aquatic ecosystems, the levels of dissolved iron are so low and some of the chemical species so unreactive that growth of cyanobacteria is impaired. Pathways of iron uptake through cyanobacterial membranes are now being elucidated, but the molecular details are still largely unknown. Here we report that the non-siderophore-producing cyanobacterium Synechocystis sp. PCC 6803 contains three exbB-exbD gene clusters that are obligatorily required for growth and are involved in iron acquisition. The three exbB-exbDs are redundant, but single and double mutants have reduced rates of iron uptake compared with wild-type cells, and the triple mutant appeared to be lethal. Short-term measurements in chemically well-defined medium show that iron uptake by Synechocystis depends on inorganic iron (Fe') concentration and ExbB-ExbD complexes are essentially required for the Fe' transport process. Although transport of iron bound to a model siderophore, ferrioxamine B, is also reduced in the exbB-exbD mutants, the rate of uptake at similar total [Fe] is about 800-fold slower than Fe', suggesting that hydroxamate siderophore iron uptake may be less ecologically relevant than free iron. These results provide the first evidence that ExbB-ExbD is involved in inorganic iron uptake and is an essential part of the iron acquisition pathway in cyanobacteria. The involvement of an ExbB-ExbD system for inorganic iron uptake may allow cyanobacteria to more tightly maintain iron homeostasis, particularly in variable environments where iron concentrations range from limiting to sufficient.