Regulation Of Photosynthesis Genes Research Articles

The Impact of the Major Endoribonucleases RNase E and RNase III and of the sRNA StsR on Photosynthesis Gene Expression in Rhodobacter sphaeroides Is Growth-Phase-Dependent.

Rhodobacter sphaeroides is a facultative phototrophic bacterium that performs aerobic respiration when oxygen is available. Only when oxygen is present at low concentrations or absent are pigment-protein complexes formed, and anoxygenic photosynthesis generates ATP. The regulation of photosynthesis genes in response to oxygen and light has been investigated for decades, with a focus on the regulation of transcription. However, many studies have also revealed the importance of regulated mRNA processing. This study analyzes the phenotypes of wild type and mutant strains and compares global RNA-seq datasets to elucidate the impact of ribonucleases and the small non-coding RNA StsR on photosynthesis gene expression in Rhodobacter. Most importantly, the results demonstrate that, in particular, the role of ribonuclease E in photosynthesis gene expression is strongly dependent on growth phase.

A Complex Network of Sigma Factors and sRNA StsR Regulates Stress Responses in R. sphaeroides

Adaptation of bacteria to a changing environment is often accompanied by remodeling of the transcriptome. In the facultative phototroph Rhodobacter sphaeroides the alternative sigma factors RpoE, RpoHI and RpoHII play an important role in a variety of stress responses, including heat, oxidative stress and nutrient limitation. Photooxidative stress caused by the simultaneous presence of chlorophylls, light and oxygen is a special challenge for phototrophic organisms. Like alternative sigma factors, several non-coding sRNAs have important roles in the defense against photooxidative stress. RNAseq-based transcriptome data pointed to an influence of the stationary phase-induced StsR sRNA on levels of mRNAs and sRNAs with a role in the photooxidative stress response. Furthermore, StsR also affects expression of photosynthesis genes and of genes for regulators of photosynthesis genes. In vivo and in vitro interaction studies revealed that StsR, that is under control of the RpoHI and RpoHII sigma factors, targets rpoE mRNA and affects its abundance by altering its stability. RpoE regulates expression of the rpoHII gene and, consequently, expression of stsR. These data provide new insights into a complex regulatory network of protein regulators and sRNAs involved in defense against photooxidative stress and the regulation of photosynthesis genes.

Contribution of Hfq to photooxidative stress resistance and global regulation in Rhodobacter sphaeroides

The photosynthetic alphaproteobacterium Rhodobacter sphaeroides has to cope with photooxidative stress that is caused by the bacteriochlorophyll a-mediated formation of singlet oxygen ((1)O(2)). Exposure to (1)O(2) induces the alternative sigma factors RpoE and RpoH(II) which then promote transcription of photooxidative stress-related genes, including small RNAs (sRNAs). The ubiquitous RNA chaperone Hfq is well established to interact with and facilitate the base-pairing of sRNAs and target mRNAs to influence mRNA stability and/or translation. Here we report on the pleiotropic phenotype of a Δhfq mutant of R. sphaeroides, which is less pigmented, produces minicells and is more sensitive to (1)O(2). The higher (1)O(2) sensitivity of the Δhfq mutant is paralleled by a reduced RpoE activity and a disordered induction of RpoH(II)-dependent genes. We used co-immunoprecipitation of FLAG-tagged Hfq combined with RNA-seq to identify association of at least 25 sRNAs and of mRNAs encoding cell division proteins and ribosomal proteins with Hfq. Remarkably, > 70% of the Hfq-bound sRNAs are (1)O(2)-affected. Proteomics analysis of the Hfq-deficient strain revealed an impact of Hfq on amino acid transport and metabolic functions. Our data demonstrate for the first time an involvement of Hfq in regulation of photosynthesis genes and in the photooxidative stress response.

A cryptochrome‐like protein is involved in the regulation of photosynthesis genes in Rhodobacter sphaeroides

Blue light receptors belonging to the cryptochrome/photolyase family are found in all kingdoms of life. The functions of photolyases in repair of UV-damaged DNA as well as of cryptochromes in the light-dependent regulation of photomorphogenetic processes and in the circadian clock in plants and animals are well analysed. In prokaryotes, the only role of members of this protein family that could be demonstrated is DNA repair. Recently, we identified a gene for a cryptochrome-like protein (CryB) in the alpha-proteobacterium Rhodobacter sphaeroides. The protein lacks the typical C-terminal extension of cryptochromes, and is not related to the Cry DASH family. Here we demonstrate that CryB binds flavin adenine dinucleotide that can be photoreduced by blue light. CryB binds single-stranded DNA with very high affinity (K(d) approximately 10(-8) M) but double-stranded DNA and single-stranded RNA with far lower affinity (K(d) approximately 10(-6) M). Despite of that, no in vitro repair activity for pyrimidine dimers in single-stranded DNA could be detected. However, we show that CryB clearly affects the expression of genes for pigment-binding proteins and consequently the amount of photosynthetic complexes in R. sphaeroides. Thus, for the first time a role of a bacterial cryptochrome in gene regulation together with a biological function is demonstrated.

Rhodobacter sphaeroides에서의 광합성유전자(puf, puc, puhA, bchC, bchE, bchF와 bchI)의 발현조절

본 연구에서는 lacZ transcriptional fusion plasmid를 이용하여 광합성 세균인 Rhodobacter sphaeroides에서의 7가지 광합성유전자 (puf, puc, puhA, bchC, bchE, bchF, bchI) 발현의 경향과 조절을 조사하였다. R. sphaeroides에서 puhA와 bchI를 제외한 모든 광합성유전자들이 호기적 조건과 비교했을 때 혐기적 조건에서 더욱 강하게 발현되었다. puhA 유전자는 bchFNBHLM-RSP0290과 operon을 형성하며, bchI 유전자는 crtA와 operon을 이루는 것으로 나타났다. 광합성 조건에서 자란 R. sphaeroides의 puf, puc, bchCXYZ operon의 발현은 빛의 세기에 비례하는 반면, bchFNBHLM(RSP0290 puhA) operon의 발현은 빛의 세기에 반비례 하였다. bchEJG의 발현은 <TEX>$10\;W/m^2$</TEX>의 빛이 조사된 광합성 조건에서 제일 낮았으며, <TEX>$100\;W/m^2$</TEX>의 빛의 광합성 조건에서 가장 높았다. R. sphaeroides의 산소인지와 빛 인지에 관련된 세 가지 주요 조절기작에 의한 광합성유전자 조절은 다음과 같다. puf와 bchC는 PpsR repressor와 PrrBA two-component system에 의해 조절된다. 그리고 puc operon은 PpsR, FnrL, PrrBA system에 의해 조절된다. bchE의 발현은 FnrL과 PrrBA system에 의해 조절되는 반면, bchF는 오로지 PpsR에 의해서만 조절된다. PpsR repressor는 강한 세기의 빛 조건에서 bchf 발현억제의 원인이 되며, FnrL은 그 자체가 산소를 인지하는 기능 이외에도 세포질의 산화/환원 상태의 인지에 관련될 것으로 보인다. Here we examined the expression patterns and regulation of seven photosynthesis (PS) genes (puf, puc, puhA, bchC, bchE, bchF, and bchI) in the anoxygenic photosynthetic bacterium, Rhodobacter sphaeroides, based on lacZ reporter gene assay. Expression of the tested PS genes, except puhA and bchI, were strongly induced in R. sphaeroides grown under anaerobic conditions relative to that under aerobic conditions. The puhA and bchI genes appear to form the operons together with bchFNBHLM-RSP0290 and crtA, respectively. Expression of the puf, puc, and bchCXYZ operons in R. sphaeroides grown photosynthetically was proportional to the incident light intensity, whereas that of bchFNBHLM(RSP0290-puhA) was inversely related to light intensity. Expression of bchEJG was lowest under medium-light photosynthetic conditions <TEX>$(10\;W/m^2)$</TEX> and highest under high light conditions <TEX>$(100\;W/m^2)$</TEX>. The regulation of PS genes by the three major regulatory systems involved in oxygen- and light-sensing in R. sphaeroides is as following: puf and bchC are regulated by both the PpsR repressor and the PrrBA two-component system. The puc operon is under control of PpsR, FnrL, and PrrBA system. Expression of bchE is controlled by FnrL and PrrBA two-component system, whereas bchF is regulated exclusively by PpsR. It was demonstrated that the PpsR repressor is responsible for high-light repression of bchF and that FnrL might be involved in perceiving the cellular redox state in addition to sensing <TEX>$O_2$</TEX> itself.

Light‐dependent regulation of photosynthesis genes in Rhodobacter sphaeroides 2.4.1 is coordinately controlled by photosynthetic electron transport via the PrrBA two‐component system and the photoreceptor AppA

Formation of the photosynthetic apparatus in Rhodobacter is regulated by oxygen tension and light intensity. Here we show that in anaerobically grown Rhodobacter cells a light-dependent increase in expression of the puc and puf operons encoding structural proteins of the photosynthetic complexes requires an active photosynthetic electron transport. The redox-sensitive CrtJ/PpsR repressor of photosynthesis genes, which was suggested to mediate electron transport-dependent signals, is not involved in this light-dependent signal chain. Our data reveal that the signal initiated in the photosynthetic reaction centre is transmitted via components of the electron transport chain and the PrrB/PrrA two-component system in Rhodobacter sphaeroides. Under blue light illumination in the absence of oxygen this signal leads to activation of photosynthesis genes and interferes with a blue-light repression mediated by the AppA photoreceptor and the PpsR transcriptional repressor in R. sphaeroides. Thus, light either sensed by a photoreceptor or initiating photosynthetic electron transport has opposite effects on the transcription of photosynthesis genes. Both signalling pathways involve redox-dependent steps that finally determine the effect of light on gene expression.

Regulation of photosynthesis genes in Rubrivivax gelatinosus: transcription factor PpsR is involved in both negative and positive control.

Induction of biosynthesis of the photosystem in anoxygenic photosynthetic bacteria occurs when the oxygen concentration drops. Control of this induction takes place primarily at the transcriptional level, with photosynthesis genes expressed preferentially under anaerobic conditions. Here, we report analysis of the transcriptional control of two photosynthesis promoters, pucBA and crtI, by the PpsR factor in Rubrivivax gelatinosus. This was accomplished by analyzing the photosystem production in the wild type and in the PPSRK (ppsR::Km) mutant grown under anaerobic and semiaerobic conditions and by assessing the beta-galactosidase activity of lacZ transcriptionally fused to promoters possessing the putative PpsR-binding consensus sequences. It was found that under semiaerobic conditions, inactivation of the ppsR gene resulted in overproduction of carotenoid and bacteriochlorophyll pigments, while the production of LH2 was drastically reduced. The beta-galactosidase activity showed that, in contrast to what has been found previously for Rhodobacter species, PpsR acts in R. gelatinosus as an aerobic repressor of the crtI gene while it acts as an activator for the expression of pucBA. Inspection of the putative PpsR-binding consensus sequences revealed significant differences that may explain the different levels of expression of the two genes studied.

Coregulation of light-harvesting complex II phosphorylation and lhcb mRNA accumulation in winter rye.

Winter rye plants grown under contrasting environmental conditions or just transiently shifted to varying light and temperature conditions, were studied to elucidate the chloroplast signal involved in regulation of photosynthesis genes in the nucleus. Photosystem II excitation pressure, reflecting the plastoquinone redox state, and the phosphorylation level of thylakoid light-harvesting proteins (LHCII and CP29) were monitored together with changes occurring in the accumulation of lhcb, rbcS and psbA mRNAs. Short-term shifts of plants to changed conditions, from 1 h to 2 d, were postulated to reveal signals crucial for the initiation of the acclimation process. Comparison of these results with those obtained from plants acclimated during several weeks' growth at contrasting temperature and in different light regimes, allow us to make the following conclusions: (1) LHCII protein phosphoylation is a sensitive tool to monitor redox changes in chloroplasts; (2) LHCII protein phosphorylation and lhcb mRNA accumulation occur under similar conditions and are possibly coregulated via an activation state of the same kinase (the LHCII kinase); (3) Maximal accumulation of lhcb mRNA during the diurnal light phase seems to require an active LHCII kinase whereas inactivation of the kinase is accompanied by dampening of the circadian oscillation in the amount of lhcb mRNA; (4) Excitation pressure of photosystem II (reduction state of the plastoquinone pool) is not directly involved in the regulation of lhcb mRNA accumulation. Instead (5) the redox status of the electron acceptors of photosystem I in the stromal compartment seems to be highly regulated and crucial for the regulation of lhcb gene expression in the nucleus.

Influence of light on the regulation of photosynthesis genes in Rhodobacter spec.

Facultatively phototrophic purple bacteria regulate the formation of their photosynthetic apparatus in response to oxygen tension and light intensity. A dropping oxygen tension increases the formation of the photosynthetic apparatus even in the absence of light. Although light is not the external stimulus that determines whether photosynthetic complexes are synthesized, the light intensity does affect the expression of photosynthesis genes in Rhodobacter. The amount of photosynthetic complexes is higher under low light than under high light intensities during anaerobic growth. The regulation of genes specific for anoxygenic photosynthesis has been studied most extensively in the related strains Rhodobacter capsulatus and Rhodobacter sphaeroides. Genes coding for the pigment binding proteins of both strains are organized in the polycistronic puf and puc operons respectively. At low oxygen tension blue light (420 nm) is most efficient in repressing R. sphaeroides puf and puc expression (1). By using Northern blot analysis we revealed that in contrast to R. sphaeroides blue light has no effect on R. capsulatus puf and puc mRNA levels during semi-aerobic growth. Furthermore we present evidence that the R sphaeroides AppA protein, capable of FAD binding (2), is involved in this light dependent gene regulation. We currently investigate the mechanisms of signal transduction and the function of AppA as potential light receptor.

Redox regulation of PSI nuclear genes through photosynthetic electron flow

Light is one of the most important environmental cues that regulate photosynthesis genes. Recently, significance of the redox regulatory system has been suggested in the light regulation of photosynthesis genes, in cyanobacteria, eukaryotic algae, and also in higher plants. In this study, we examined how PSI nuclear genes of Chlamydomonas reinhardtii are regulated in response to light. When Chlamydomonas cells were transferred from the darkness to the light, psaE mRNA level was drastically increased. This light induction was inhibited by the photosynthetic electron transport inhibitors, DCMU and DBMIB, but not by an uncoupler, CCCP. These demonstrate that, in this induction, photosynthetic electron flow plays a pivotal role but photophosphorylation is not necessary. Subsequently, we examined whether redox state of intersystem electron carrier is responsible for this induction. For this sake, we irradiated the Chlamydomonas cells with PSI-light (695nm) or PSII-light (644nm), which makes plastoquinone pool oxidative and reductive, respectively. However, both monochromes induced the psaE mRNA to similar extent, indicating that the redox state of intersystem electron carriers, e.g. plastoquinone pool, is not responsible for this light induction. We are currently examining how the photosynthesis-derived signal causes this response, and several hints obtained by our recent experiments will be discussed.

Structure and regulation of photosynthesis genes in Pinus sylvestris (Scots pine) and Pinus contorta (lodgepole pine)

The structure and regulation of one nuclear and one chloroplast gene was studied in Pinus sylvestris (Scots pine) and Pinus contorta (lodgepole pine). cDNA copies of the nuclear located cab genes of Pinus sylvestris, coding for the light-harvesting chlorophyll a/b-binding proteins of photosystem II (LHC-II), were cloned. cab-II genes coding for both types of LHC-II polypeptides, Types 1 and 2, were found. An analysis of the DNA sequences of several different cab-II cDNAs shows that they have a high bias for the nucleotides G and C at the third base positions of the codons, making them more similar to monocot than to dicot genes. Two of the three genes were found to be located within CpG islands. The cab-II genes were found to be expressed in dark-grown seedlings in contrast to what has been found for most angiosperms. The chloroplast genomes of conifers were shown to lack the inverted repeat organization normally found in higher plants, mosses and green algae. The psbA gene, located in the chloroplast genome and coding for the D1 polypeptide in the reaction center of photosystem II, wasfound to be tandemly duplicated in P. contorta. Cloning and sequence analysis of the two psbA genes and the surrounding regions showed that the duplicated segment is 1.97 kb long and that it ends 19 bp downstream from the psbA stop codon. The corresponding locus of P. sylvestris, which lacks the duplication, was cloned and characterized. A comparison with P. contorta indicates how the duplication/insertion event has occurred. A comparison of third codon position between P. contorta psbA and that of other plants indicated an almost equidistant evolutionary relationship between P. contorta, spinach (or barley) and Marchantia polymorpha.