Photosystem I Genes Research Articles

Physcomitrium patens pentatricopeptide repeat protein PpPPR_32 is involved in the accumulation of psaC mRNA encoding the iron sulfur protein of photosystem I.

Pentatricopeptide repeat (PPR) proteins are involved in RNA metabolism and also play a role in posttranscriptional regulation during plant organellar gene expression. Although a hundred of PPR proteins exist in the moss Physcomitrium patens, their functions are not fully understood. Here, we report the function of P-class PPR protein PpPPR_32 in P. patens. A transient expression assay using green fluorescent protein demonstrated that the N-terminal region of PpPPR_32 functions as a chloroplast-targeting transit peptide, indicating that PpPPR_32 is localized in chloroplasts. PpPPR_32 knockout mutants grew autotrophically but with reduced protonema growth and the poor formation of photosystem I (PSI) complexes. Quantitative real-time reverse transcription-polymerase chain reaction and RNA gel blot hybridization analyses revealed a significant reduction in the transcript level of the psaC gene encoding the iron sulfur protein of PSI but no alteration to the transcript levels of other PSI genes. This suggests that PpPPR_32 is specifically involved in the expression level of the psaC gene. Our results indicate that PpPPR_32 is essential for the accumulation of psaC transcript and PSI complexes.

Exogenous melatonin alleviates NO2 damage in tobacco leaves by promoting antioxidant defense, modulating redox homeostasis, and signal transduction

Nitrogen dioxide (NO2) is a common outdoor air pollutant, which has adverse effects on the environment and human health. Herein, NO2 inhibited photosynthesis and antioxidant capacity in plants. Melatonin (Mel) is a neurohormone found in the pineal gland. Exogenous Mel alleviated chlorophyll degradation and increased the expression of key proteins and genes in the process of chlorophyll synthesis in tobacco leaves exposed to NO2. Additionally, the activities of photosystem II (PSII) and photosystem I (PSI) were enhanced. PSII and PSI reaction center proteins and genes were upregulated. Mel pre-treatment enhanced enzyme activities and expression of proteins related to the ascorbic acid–glutathione cycle and thioredoxin-peroxiredoxin pathway in leaves exposed to NO2, thus regulating their redox balance. Furthermore, exogenous Mel mediated the polyamine synthesis pathway and increased the expression of the key enzyme proteins SAMS1, SAMS2, and SAMS3 in the polyamine synthesis pathway in leaves under NO2 stress. Mel regulated ABA signal transduction and calmodulin binding transcription factors CAMTA12 and NtCaM calmodulin NtCaM2 in Ca2+ signal transduction. Collectively, these results elucidate that Mel can alleviate high-concentration NO2, thus suitable for agricultural application.

Structural basis for the adaptation and function of chlorophyll f in photosystem I

Chlorophylls (Chl) play pivotal roles in energy capture, transfer and charge separation in photosynthesis. Among Chls functioning in oxygenic photosynthesis, Chl f is the most red-shifted type first found in a cyanobacterium Halomicronema hongdechloris. The location and function of Chl f in photosystems are not clear. Here we analyzed the high-resolution structures of photosystem I (PSI) core from H. hongdechloris grown under white or far-red light by cryo-electron microscopy. The structure showed that, far-red PSI binds 83 Chl a and 7 Chl f, and Chl f are associated at the periphery of PSI but not in the electron transfer chain. The appearance of Chl f is well correlated with the expression of PSI genes induced under far-red light. These results indicate that Chl f functions to harvest the far-red light and enhance uphill energy transfer, and changes in the gene sequences are essential for the binding of Chl f.

Diversity of viral photosystem-I psaA genes.

Marine photosynthesis is one of the major contributors to the global carbon cycle and the world's oxygen supply. This process is largely driven by cyanobacteria, namely Synechococcus and Prochlorococcus. Genes encoding photosystem-II (PSII) reaction center proteins are found in many cyanophage genomes, and are expressed during the infection of their hosts. On the basis of metagenomics, cyanophage photosystem-I (PSI) gene cassettes were recently discovered with two gene arrangements psaJF→C→A→B→K→E→D and psaD→C→A→B. It was suggested that the horizontal transfer of PSII and PSI genes is increasing phage fitness. To better understand their diversity, we designed degenerate primers to cover a wide diversity of organisms, and using PCR we targeted the psaC→A arrangement, which is unique to cyanophages cassettes. We examined viral concentrates from four islands in the Pacific Ocean and found samples containing the psaC→A arrangement. Analyses of the amplified viral psaA gene revealed six subgroups varying in their level of similarity and %G+C content, suggesting that the diversity of cyanophage PSI genes is greater than originally thought.

PSBP-DOMAIN PROTEIN1, a Nuclear-Encoded Thylakoid Lumenal Protein, Is Essential for Photosystem I Assembly in Arabidopsis

To gain insights into the molecular details of photosystem I (PSI) biogenesis, we characterized the PsbP-domain protein1 (ppd1) mutant of Arabidopsis thaliana that specifically lacks PSI activity. Deletion of PPD1 results in an inability of the mutant to grow photoautotrophically and a specific loss of the stable PSI complex. Unaltered transcription and translation of plastid-encoded PSI genes indicate that PPD1 acts at the posttranslational level. In vivo protein labeling experiments reveal that the rate of synthesis of PSI reaction center proteins PsaA/B in ppd1 is comparable to that of wild-type plants, whereas the rate of turnover of PsaA/B proteins is higher in ppd1 than in wild-type plants. With increasing leaf age, PPD1 content decreases considerably, while PSI content remains constant. PPD1 is a nuclear-encoded thylakoid lumenal protein and is associated with PSI but is not an integral subunit of PSI. Biochemical and molecular analyses reveal that PPD1 interacts directly and specifically with PsaB and PsaA. Yeast two-hybrid experiments show that PPD1 interacts with some lumenal loops of PsaB and PsaA. Our results suggest that PPD1 is a PSI assembly factor that assists the proper folding and integration of PsaB and PsaA into the thylakoid membrane.

The Plastid Genome-Encoded Ycf4 Protein Functions as a Nonessential Assembly Factor for Photosystem I in Higher Plants

Photosystem biogenesis in the thylakoid membrane is a highly complicated process that requires the coordinated assembly of nucleus-encoded and chloroplast-encoded protein subunits as well as the insertion of hundreds of cofactors, such as chromophores (chlorophylls, carotenoids) and iron-sulfur clusters. The molecular details of the assembly process and the identity and functions of the auxiliary factors involved in it are only poorly understood. In this work, we have characterized the chloroplast genome-encoded ycf4 (for hypothetical chloroplast reading frame no. 4) gene, previously shown to encode a protein involved in photosystem I (PSI) biogenesis in the unicellular green alga Chlamydomonas reinhardtii. Using stable transformation of the chloroplast genome, we have generated ycf4 knockout plants in the higher plant tobacco (Nicotiana tabacum). Although these mutants are severely affected in their photosynthetic performance, they are capable of photoautotrophic growth, demonstrating that, different from Chlamydomonas, the ycf4 gene product is not essential for photosynthesis. We further show that ycf4 knockout plants are specifically deficient in PSI accumulation. Unaltered expression of plastid-encoded PSI genes and biochemical analyses suggest a posttranslational action of the Ycf4 protein in the PSI assembly process. With increasing leaf age, the contents of Ycf4 and Y3IP1, another auxiliary factor involved in PSI assembly, decrease strongly, whereas PSI contents remain constant, suggesting that PSI is highly stable and that its biogenesis is restricted to young leaves.

Viral clones from the GOS expedition with an unusual photosystem-I gene cassette organization

Cyanobacteria have a key role in marine photosynthesis, which contributes to the global carbon cycle and to the world oxygen supply. Genes encoding for photosystem-II (PSII) and photosystem-I (PSI) reaction centers are found in different cyanophage genomes, and it was suggested that the horizontal transfer of these genes might be involved in increasing phage fitness. We have further analyzed a rare viral Global Ocean Sampling (GOS) clone containing PSI genes. This clone contains the unusual PSI gene organization psaD->C->A, as opposed to the more frequently observed viral psaJF->C->A->B->K->E->D organization, and was detected only once in the GOS metagenome. Our analyses identified more occurrences with similar arrangement and indicate that this PSI viral gene organization (now psaD->C->A->B), although rare, is authentic and represents a new PSI gene arrangement.

Time‐series resolution of gradual nitrogen starvation and its impact on photosynthesis in the cyanobacterium Synechocystis PCC 6803

Sequential adaptation to nitrogen deprivation and ultimately to full starvation requires coordinated adjustment of cellular functions. We investigated changes in gene expression and cell physiology of the cyanobacterium Synechocystis PCC 6803 during 96 h of nitrogen starvation. During the first 6 h, the transcriptome showed activation of nitrogen uptake and assimilation systems and of the core nitrogen and carbon assimilation regulators. However, the nitrogen-deprived cells still grew at the same rate as the control and even showed transiently increased expression of phycobilisome genes. After 12 h, cell growth decreased and chlorosis started with degradation of the nitrogen-rich phycobilisomes. During this phase, the transcriptome showed suppression of genes for phycobilisomes, for carbon fixation and for de novo protein synthesis. Interestingly, photosynthetic activity of both photosystem I (PSI) and photosystem II was retained quite well. Excess electrons were quenched by the induction of terminal oxidase and hydrogenase genes, compensating for the diminished carbon fixation and nitrate reduction activity. After 48 h, the cells ceased most activities. A marked exception was the retained PSI gene transcription, possibly this supports the viability of Synechocystis cells and enables rapid recovery after relieving from nitrogen starvation. During early recovery, many genes changed expression, supporting the resumed cellular activity. In total, our results distinguished three phases during gradual nitrogen depletion: (1) an immediate response, (2) short-term acclimation and (3) long-term survival. This shows that cyanobacteria respond to nitrogen starvation by a cascade of physiological adaptations reflected by numerous changes in the transcriptome unfolding at different timescales.

Marine cyanophages: tinkering with the electron transport chain

Marine cyanophages: tinkering with the electron transport chain

Reconstructing a puzzle: existence of cyanophages containing both photosystem‐I and photosystem‐II gene suites inferred from oceanic metagenomic datasets

Cyanobacteria play a key role in marine photosynthesis, which contributes to the global carbon cycle and to the world oxygen supply. Genes encoding the photosystem-II (PSII) reaction centre are found in many cyanophage genomes, and it was suggested that the horizontal transfer of these genes might be involved in increasing phage fitness. Recently, evidence for the existence of phages carrying Photosystem-I (PSI) genes was also reported. Here, using a combination of different marine metagenomic datasets and a unique crossing of the datasets, we now describe the finding of phages that, as in plants and cyanobacteria, contain both PSII and PSI genes. In addition, these phages also contain NADH dehydrogenase genes. The presence of modified PSII and PSI genes in the same viral entities in combination with electron transfer proteins like NAD(P)H dehydrogenase (NDH-1) strongly points to a role in perturbation of the cyanobacterial host photosynthetic electron flow. We therefore suggest that, depending on the physiological condition of the infected cyanobacterial host, the viruses may use different options to maximize survival. The modified PSI may alternate between functioning with PSII in linear electron transfer and contributing to the production of both NADPH and ATP or functioning independently of PSII in cyclic mode via the NDH-1 complex and thus producing only ATP.

Role of multiple HLR1 sequences in the regulation of the dual promoters of the psaAB genes in Synechocystis sp. PCC 6803.

Previously, we analyzed the promoter architecture of the psaAB genes encoding reaction center subunits of photosystem I (PSI) in the cyanobacterium Synechocystis sp. PCC 6803. There exist two promoters, P1 and P2, both of which show typical high-light (HL) response of PSI genes; their activities are high under low-light (LL) conditions but rapidly downregulated upon the shift to HL conditions. In this study, it was suggested that a response regulator RpaB binds to multiple high-light regulatory 1 (HLR1) sequences in the upstream region of the psaAB genes. We explored the regulatory role of cis-elements, including these HLR1 sequences on the individual activity of P1 and P2. Under LL conditions, the most influential cis-element is HLR1C (-62 to -45, relative to the transcriptional starting point of P1) working for positive regulation of P1. The other HLR1 sequences also affect the promoter activity under LL conditions; HLR1A (-255 to -238) is involved in repression of P1, whereas HLR1B (-153 to -126) works for activation of P2. Upon the shift to HL conditions, regulation via HNE2 located within the region from -271 to -177 becomes active in order to downregulate both P1 and P2 activities. A positive effect of HLR1B on P2 may persist under HL. These results suggest that cis-elements, including multiple HLR1 sequences, differently regulate the activities of dual promoters of the psaAB genes to achieve the fine-tuning of the gene expression.

Photosystem I gene cassettes are present in marine virus genomes

Cyanobacteria of the Synechococcus and Prochlorococcus genera are important contributors to photosynthetic productivity in the open oceans. Recently, core photosystem II (PSII) genes were identified in cyanophages and proposed to function in photosynthesis and in increasing viral fitness by supplementing the host production of these proteins. Here we show evidence for the presence of photosystem I (PSI) genes in the genomes of viruses that infect these marine cyanobacteria, using pre-existing metagenomic data from the global ocean sampling expedition as well as from viral biomes. The seven cyanobacterial core PSI genes identified in this study, psaA, B, C, D, E, K and a unique J and F fusion, form a cluster in cyanophage genomes, suggestive of selection for a distinct function in the virus life cycle. The existence of this PSI cluster was confirmed with overlapping and long polymerase chain reaction on environmental DNA from the Northern Line Islands. Potentially, the seven proteins encoded by the viral genes are sufficient to form an intact monomeric PSI complex. Projection of viral predicted peptides on the cyanobacterial PSI crystal structure suggested that the viral-PSI components might provide a unique way of funnelling reducing power from respiratory and other electron transfer chains to the PSI.

The response regulator RpaB binds to the upstream element of photosystem I genes to work for positive regulation under low-light conditions in Synechocystis sp. Strain PCC 6803.

The coordinated high-light response of genes encoding subunits of photosystem I (PSI) is achieved by the AT-rich region located just upstream of the core promoter in Synechocystis sp. strain PCC 6803. The upstream element enhances the basal promoter activity under low-light conditions, whereas this positive regulation is lost immediately after the shift to high-light conditions. In this study, we focused on a high-light regulatory 1 (HLR1) sequence included in the upstream element of every PSI gene examined. A gel mobility shift assay revealed that a response regulator RpaB binds to the HLR1 sequence in PSI promoters. Base substitution in the HLR1 sequence or decrease in copy number of the rpaB gene resulted in decrease in the promoter activity of PSI genes under low-light conditions. These observations suggest that RpaB acts as a transcriptional activator for PSI genes. It is likely that RpaB binds to the HLR1 sequence under low-light conditions and works for positive regulation of PSI genes and for negative regulation of high-light-inducible genes depending on the location of the HLR1 sequence within target promoters.

Transcriptional regulation of genes encoding subunits of photosystem I during acclimation to high-light conditions in Synechocystis sp. PCC 6803.

Cyanobacteria, algae and plants reduce photosystem contents and modulate photosystem stoichiometry upon acclimation to high-light (HL) conditions to avoid the damage due to excess light energy. In order to elucidate the molecular mechanism of HL acclimation, mRNA levels of genes encoding subunits of photosystems were examined in response to change of photon flux density in the wild-type strain of Synechocystis sp. PCC 6803. Transcript levels and promoter activities of photosystem I (PSI) genes rapidly decreased upon the shift to HL to less than 10% of the initial level within 1 h, whereas responses of photosystem II (PSII) transcript levels were not coordinated. The prompt change in promoter activities of PSI genes, but not PSII genes, seems important for Synechocystis cells to regulate their photosystem contents in response to changes in photon flux density. The mRNA stabilities of PSI genes also decreased during HL incubation. The down-regulation of PSI transcripts under HL did not depend on de novo protein synthesis, contrasting with the requirement of newly synthesized protein factor(s) for the accumulation of PSI transcripts under low light. When gene expression profiles of wild-type cells and a pmgA-disrupted mutant that cannot modulate photosystem stoichiometry under HL were compared, a large increase in the psaAB and psaA transcript levels was observed in the mutant under prolonged exposure to HL for 6 h. Repression of the promoter activity of the psaA gene by a pmgA-dependent mechanism was shown to be essential for the adjustment of photosystem stoichiometry under HL conditions.

A Novel Mechanism of Nuclear Photosynthesis Gene Regulation by Redox Signals from the Chloroplast during Photosystem Stoichiometry Adjustment

Photosynthetic organisms acclimate to long term changes in the environmental light quality by an adjustment of their photosystem stoichiometry to maintain photosynthetic efficiency. By using light sources that predominantly excite either photosystem I (PSI) or photosystem II (PSII), we studied the effects of excitation imbalances between both photosystems on nuclear PSI gene transcription in transgenic tobacco seedlings with promoter::beta-glucuronidase gene fusions. Shifts from PSI to PSII light sources (and vice versa) induced changes in the reduction/oxidation state of intersystem redox components, and acclimation of tobacco seedlings to such changes were monitored by changes in chlorophyll a/b ratios and in vivo chlorophyll a fluorescence. The ferredoxin-NADP(+)-oxidoreductase gene promoter did not respond to these treatments, those from the genes for subunits PsaD and PsaF of PSI are activated by a reduction signal, and the plastocyanin promoter responded to both reduction and oxidation signals. Additional experiments with photosynthetic electron transport inhibitors 3-(3',4'-dichlorophenyl)-1,1'-dimethyl urea and 2,5-dibromo-3-methyl-6-isopropyl-p-benzoquinone demonstrated that the redox state of the plastoquinone pool controls the activity of the plastocyanin promoter, whereas subunit PsaD and PsaF gene transcription is regulated by other photosynthesis-derived signals. Thus, the expression of nuclear-encoded PSI genes is controlled by diverse light quality-dependent redox signals from the plastids during photosystem stoichiometry adjustment.

Tobacco Nuclear Genes for Photosystem I Subunits, <italic>psaD, psaE</italic> and <italic>psaH</italic> Share an Octamer Motif Bound with Nuclear Proteins

In Nicotiana sylvestris, nuclear-encoded photosystem I (PSI) genes, psaD, psaE and psaH, share an octamer motif bound with three phosphoproteins. This motif is not found in the chloroplast genome. From the view point of endosymbiont hypothesis, these results suggest that a set of ancient PSI genes acquired a common cis-element in the nucleus after they were transferred from the ancestral organelle.

Differential expression inArabidopsis ofLhca2, a PSI cab gene

A cDNA clone of the gene Lhca2 encoding a photosystem I (PSI) type II chlorophyll a/b-binding protein was isolated from Arabidopsis thaliana. The isolation of this, the fourth PSI cab gene from Arabidopsis, confirms a previous report [1] that indicated Arabidopsis may contain all four PSI cab genes identified in other plant species. Lhca2 is a single-copy gene as are the other known Arabidopsis PSI cab genes. The patterns of developmental expression and tissue-specific regulation of Lhca2 are similar to those of other PSI and PSII cab genes, but the light induction pattern and the steady-state mRNA level of Lhca2 are distinct. This suggests that a different mechanism may be employed to regulate the expression of Lhca2.

Light quality regulates expression of chloroplast genes and assembly of photosynthetic membrane complexes

The concentrations of photosystem I (PSI) and photosystem II (PSII) reaction centers and the level of chloroplast reaction center gene transcripts were determined in pea plants grown under different light-quality regimes. In plants grown in light primarily absorbed by PSI ("red" light), the PSII/PSI reaction center ratio was 2-fold greater than that in plants grown in PSII-sensitizing ("yellow") light. In addition, the ratio of a PSII gene (psbB) transcript to a PSI gene (psaA) transcript was 2.6 times greater in red-grown plants relative to yellow-grown plants. Thus, a differential reaction-center concentration in the thylakoid membrane was accompanied by a differential expression of reaction center genes, suggesting that the synthesis of chloroplast membrane complexes and the assembly of photosystems are regulated by light quality at the transcriptional and/or post-transcriptional level.