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1.
Physiol Plant ; 176(2): e14227, 2024.
Artículo en Inglés | MEDLINE | ID: mdl-38410876

RESUMEN

In addition to leaves, the main site of photosynthetic reactions, active photosynthesis also takes place in stems, siliques and tree trunks. Although non-foliar photosynthesis has a marked effect on plant growth and yield, only limited information on the expression patterns of photosynthesis-related genes and the structure of photosynthetic machinery in different plant organs has been available. Here, we report the results of transcriptomic analysis of various organs of Arabidopsis thaliana and compare the gene expression profiles of young and mature leaves with a special focus on photosynthetic genes. Further, we analyzed the composition and organization of the photosynthetic electron transfer machinery in leaves, stems and green siliques at the protein level using BN-PAGE. RNA-Seq analysis revealed unique gene expression profiles in different plant organs and showed major differences in the expression of photosynthesis-related genes in young as compared to mature rosettes. Gel-based proteomic analysis of the thylakoid protein complex organization further showed that all studied plant organs contain the necessary components of the photosynthetic electron transfer chain. Intriguingly, stems accumulate high amounts of PSI-NDH complex, which has previously been implicated in cyclic electron transfer.


Asunto(s)
Proteínas de Arabidopsis , Arabidopsis , Arabidopsis/genética , Arabidopsis/metabolismo , Transcriptoma , Proteómica , Fotosíntesis/genética , Transporte de Electrón/genética , Proteínas de Arabidopsis/genética , Proteínas de Arabidopsis/metabolismo , Plantas/metabolismo , Perfilación de la Expresión Génica , Complejo de Proteína del Fotosistema I/metabolismo
2.
Plant Cell Physiol ; 64(5): 549-563, 2023 May 15.
Artículo en Inglés | MEDLINE | ID: mdl-37026998

RESUMEN

Acetylation is one of the most common chemical modifications found on a variety of molecules ranging from metabolites to proteins. Although numerous chloroplast proteins have been shown to be acetylated, the role of acetylation in the regulation of chloroplast functions has remained mainly enigmatic. The chloroplast acetylation machinery in Arabidopsis thaliana consists of eight General control non-repressible 5 (GCN5)-related N-acetyltransferase (GNAT)-family enzymes that catalyze both N-terminal and lysine acetylation of proteins. Additionally, two plastid GNATs have also been reported to be involved in the biosynthesis of melatonin. Here, we have characterized six plastid GNATs (GNAT1, GNAT2, GNAT4, GNAT6, GNAT7 and GNAT10) using a reverse genetics approach with an emphasis on the metabolomes and photosynthesis of the knock-out plants. Our results reveal the impact of GNAT enzymes on the accumulation of chloroplast-related compounds, such as oxylipins and ascorbate, and the GNAT enzymes also affect the accumulation of amino acids and their derivatives. Specifically, the amount of acetylated arginine and proline was significantly decreased in the gnat2 and gnat7 mutants, respectively, as compared to the wild-type Col-0 plants. Additionally, our results show that the loss of the GNAT enzymes results in increased accumulation of Rubisco and Rubisco activase (RCA) at the thylakoids. Nevertheless, the reallocation of Rubisco and RCA did not have consequent effects on carbon assimilation under the studied conditions. Taken together, our results show that chloroplast GNATs affect diverse aspects of plant metabolism and pave way for future research into the role of protein acetylation.


Asunto(s)
Arabidopsis , Arabidopsis/metabolismo , Acetiltransferasas/metabolismo , Ribulosa-Bifosfato Carboxilasa/metabolismo , Cloroplastos/metabolismo , Fotosíntesis
3.
Physiol Plant ; 175(5): e13999, 2023.
Artículo en Inglés | MEDLINE | ID: mdl-37882278

RESUMEN

Disassembly and degradation of the photosynthetic protein complexes during autumn senescence, a vital step to ensure efficient nutrient relocalization for winter storage, is poorly understood. Concomitantly with the degradation, anthocyanins are often synthesized. However, as to why leaves accumulate red pigments, no consensus exists. One possibility is that anthocyanins protect senescing leaves from excess light. In this study, we investigated the pigment composition, photosynthetic performance, radical production, and degradation of the photosynthetic protein complexes in Norway maple (Acer platanoides) and in its highly pigmented, purple-colored variety (Faassen's black) during autumn senescence, to dissect the possible roles of anthocyanins in photoprotection. Our findings show that senescing Faassen's black was indeed more resistant to Photosystem II (PSII) photoinhibition, presumably due to its high anthocyanin content, than the green maple. However, senescing Faassen's black exhibited low photosynthetic performance, probably due to a poor capacity to repair PSII. Furthermore, an analysis of photosynthetic protein complexes demonstrated that in both maple varieties, the supercomplexes consisting of PSII and its antenna were disassembled first, followed by the degradation of the PSII core, Photosystem I, Cytochrome b6 f, and ATP synthase. Strikingly, the degradation process appeared to proceed faster in Faassen's black, possibly explaining its poor PSII repair capacity. The results suggest that tolerance against PSII photoinhibition may not necessarily translate to a better fitness. Finally, thylakoids isolated from senescing and non-senescing leaves of both maple varieties accumulated very little carbon-centered radicals, suggesting that thylakoids may not be a major source of reactive oxygen species in senescing leaves.


Asunto(s)
Acer , Antocianinas , Antocianinas/metabolismo , Clorofila/metabolismo , Fotosíntesis/fisiología , Complejo de Proteína del Fotosistema II/metabolismo , Hojas de la Planta/metabolismo , Luz
4.
Plant Cell Physiol ; 63(9): 1205-1214, 2022 Sep 15.
Artículo en Inglés | MEDLINE | ID: mdl-35792507

RESUMEN

Higher plants acclimate to changes in light conditions by adjusting the thylakoid membrane ultrastructure. Additionally, excitation energy transfer between photosystem II (PSII) and photosystem I (PSI) is balanced in a process known as state transition. These modifications are mediated by reversible phosphorylation of Lhcb1 and Lhcb2 proteins in different pools of light-harvesting complex (LHCII) trimers. Our recent study demonstrated that chloroplast acetyltransferase NUCLEAR SHUTTLE INTERACTING (NSI)/GNAT2 (general control non-repressible 5 (GCN5)-related N-acetyltransferase 2) is also needed for the regulation of light harvesting, evidenced by the inability of the gnat2 mutant to perform state transitions although there are no defects in LHCII phosphorylation. Here, we show that despite contrasting phosphorylation states of LHCII, grana packing in the gnat2 and state transition 7 (stn7) mutants possesses similar features, as the thylakoid structure of the mutants does not respond to the shift from darkness to light, which is in striking contrast to wild type (Wt). Circular dichroism and native polyacrylamide gel electrophoresis analyses further revealed that the thylakoid protein complex organization of gnat2 and stn7 resembles each other, but differ from that of Wt. Also, the location of the phosphorylated Lhcb2 as well as the LHCII antenna within the thylakoid network in gnat2 mutant is different from that of Wt. In gnat2, the LHCII antenna remains largely in grana stacks, where the phosphorylated Lhcb2 is found in all LHCII trimer pools, including those associated with PSII. These results indicate that in addition to phosphorylation-mediated regulation through STN7, the GNAT2 enzyme is involved in the organization and dynamics of thylakoid structure, probably through the regulation of chloroplast protein acetylation.


Asunto(s)
Arabidopsis , Tilacoides , Acetiltransferasas/metabolismo , Arabidopsis/genética , Arabidopsis/metabolismo , Cloroplastos/metabolismo , Complejos de Proteína Captadores de Luz/metabolismo , Fosforilación , Complejo de Proteína del Fotosistema I/metabolismo , Complejo de Proteína del Fotosistema II/metabolismo , Tilacoides/metabolismo
5.
Mol Syst Biol ; 16(7): e9464, 2020 07.
Artículo en Inglés | MEDLINE | ID: mdl-32633465

RESUMEN

Protein acetylation is a highly frequent protein modification. However, comparatively little is known about its enzymatic machinery. N-α-acetylation (NTA) and ε-lysine acetylation (KA) are known to be catalyzed by distinct families of enzymes (NATs and KATs, respectively), although the possibility that the same GCN5-related N-acetyltransferase (GNAT) can perform both functions has been debated. Here, we discovered a new family of plastid-localized GNATs, which possess a dual specificity. All characterized GNAT family members display a number of unique features. Quantitative mass spectrometry analyses revealed that these enzymes exhibit both distinct KA and relaxed NTA specificities. Furthermore, inactivation of GNAT2 leads to significant NTA or KA decreases of several plastid proteins, while proteins of other compartments were unaffected. The data indicate that these enzymes have specific protein targets and likely display partly redundant selectivity, increasing the robustness of the acetylation process in vivo. In summary, this study revealed a new layer of complexity in the machinery controlling this prevalent modification and suggests that other eukaryotic GNATs may also possess these previously underappreciated broader enzymatic activities.


Asunto(s)
Arabidopsis/metabolismo , Lisina/química , Acetiltransferasas N-Terminal/metabolismo , Proteínas de Plantas/metabolismo , Plastidios/genética , Plastidios/metabolismo , Acetilación , Arabidopsis/enzimología , Arabidopsis/genética , Cloroplastos/enzimología , Cloroplastos/metabolismo , Cromatografía Líquida de Alta Presión , Cromatografía Liquida , Epigenoma , Escherichia/genética , Escherichia/metabolismo , Técnicas de Inactivación de Genes , Genoma de Planta , Técnicas In Vitro , Acetiltransferasas N-Terminal/química , Acetiltransferasas N-Terminal/genética , Péptidos/química , Péptidos/genética , Filogenia , Proteínas de Plantas/genética , Plastidios/enzimología , Proteínas Recombinantes , Espectrometría de Masas en Tándem
6.
Plant Cell Environ ; 44(2): 548-558, 2021 02.
Artículo en Inglés | MEDLINE | ID: mdl-33131061

RESUMEN

In Arabidopsis, two leaf-type ferredoxin-NADP+ oxidoreductase (LFNR) isoforms function in photosynthetic electron flow in reduction of NADP+ , while two root-type FNR (RFNR) isoforms catalyse reduction of ferredoxin in non-photosynthetic plastids. As the key to understanding, the function of RFNRs might lie in their spatial and temporal distribution in different plant tissues and cell types, we examined expression of RFNR1 and RFNR2 genes using ß-glucuronidase (GUS) reporter lines and investigated accumulation of distinct RFNR isoforms using a GFP approach and Western blotting upon various stresses. We show that while RFNR1 promoter is active in leaf veins, root tips and in the stele of roots, RFNR2 promoter activity is present in leaf tips and root stele, epidermis and cortex. RFNR1 protein accumulates as a soluble protein within the plastids of root stele cells, while RFNR2 is mainly present in the outer root layers. Ozone treatment of plants enhanced accumulation of RFNR1, whereas low temperature treatment specifically affected RFNR2 accumulation in roots. We further discuss the physiological roles of RFNR1 and RFNR2 based on characterization of rfnr1 and rfnr2 knock-out plants and show that although the function of these proteins is partly redundant, the RFNR proteins are essential for plant development and survival.


Asunto(s)
Proteínas de Arabidopsis/genética , Arabidopsis/enzimología , Ferredoxina-NADP Reductasa/metabolismo , Oxidorreductasas/genética , Arabidopsis/genética , Proteínas de Arabidopsis/metabolismo , Frío , Transporte de Electrón , Ferredoxina-NADP Reductasa/genética , Oxidorreductasas/metabolismo , Fotosíntesis , Hojas de la Planta/enzimología , Hojas de la Planta/genética , Raíces de Plantas/enzimología , Raíces de Plantas/genética , Plastidios/enzimología , Isoformas de Proteínas , Estrés Fisiológico
7.
Plant Cell ; 30(8): 1695-1709, 2018 08.
Artículo en Inglés | MEDLINE | ID: mdl-29967049

RESUMEN

The amount of light energy received by the photosynthetic reaction centers photosystem II (PSII) and photosystem I (PSI) is balanced through state transitions. Reversible phosphorylation of a light-harvesting antenna trimer (L-LHCII) orchestrates the association between L-LHCII and the photosystems, thus adjusting the amount of excitation energy received by the reaction centers. In this study, we identified the enzyme NUCLEAR SHUTTLE INTERACTING (NSI; AT1G32070) as an active lysine acetyltransferase in the chloroplasts of Arabidopsis thaliana Intriguingly, nsi knockout mutant plants were defective in state transitions, even though they had a similar LHCII phosphorylation pattern as the wild type. Accordingly, nsi plants were not able to accumulate the PSI-LHCII state transition complex, even though the LHCII docking site of PSI and the overall amounts of photosynthetic protein complexes remained unchanged. Instead, the nsi mutants showed a decreased Lys acetylation status of specific photosynthetic proteins including PSI, PSII, and LHCII subunits. Our work demonstrates that the chloroplast acetyltransferase NSI is needed for the dynamic reorganization of thylakoid protein complexes during photosynthetic state transitions.


Asunto(s)
Arabidopsis/metabolismo , Cloroplastos/enzimología , Arabidopsis/genética , Cloroplastos/genética , Mutación , Fosforilación/genética , Fosforilación/fisiología , Complejo de Proteína del Fotosistema I/metabolismo , Complejo de Proteína del Fotosistema II/metabolismo
8.
Plant J ; 99(2): 245-256, 2019 07.
Artículo en Inglés | MEDLINE | ID: mdl-30888718

RESUMEN

During daffodil flower development, chloroplasts differentiate into photosynthetically inactive chromoplasts having lost functional photosynthetic reaction centers. Chromoplasts exhibit a respiratory activity reducing oxygen to water and generating ATP. Immunoblots revealed the presence of the plastid terminal oxidase (PTOX), the NAD(P)H dehydrogenase (NDH) complex, the cytochrome b6 f complex, ATP synthase and several isoforms of ferredoxin-NADP+ oxidoreductase (FNR), and ferredoxin (Fd). Fluorescence spectroscopy allowed the detection of chlorophyll a in the cytochrome b6 f complex. Here we characterize the electron transport pathway of chromorespiration by using specific inhibitors for the NDH complex, the cytochrome b6 f complex, FNR and redox-inactive Fd in which the iron was replaced by gallium. Our data suggest an electron flow via two separate pathways, both reducing plastoquinone (PQ) and using PTOX as oxidase. The first oxidizes NADPH via FNR, Fd and cytochrome bh of the cytochrome b6 f complex, and does not result in the pumping of protons across the membrane. In the second, electron transport takes place via the NDH complex using both NADH and NADPH as electron donor. FNR and Fd are not involved in this pathway. The NDH complex is responsible for the generation of the proton gradient. We propose a model for chromorespiration that may also be relevant for the understanding of chlororespiration and for the characterization of the electron input from Fd to the cytochrome b6 f complex during cyclic electron transport in chloroplasts.


Asunto(s)
Transporte de Electrón , Narcissus/metabolismo , Plastidios/metabolismo , Clorofila A/metabolismo , Complejo de Citocromo b6f/metabolismo , Ferredoxinas/metabolismo , NADP/metabolismo , Oxidación-Reducción , Fotosíntesis , Proteínas del Complejo del Centro de Reacción Fotosintética
9.
BMC Plant Biol ; 20(1): 413, 2020 Sep 04.
Artículo en Inglés | MEDLINE | ID: mdl-32887556

RESUMEN

BACKGROUND: Non-photosynthetic plastids of plants are known to be involved in a range of metabolic and biosynthetic reactions, even if they have been difficult to study due to their small size and lack of color. The morphology of root plastids is heterogeneous and also the plastid size, density and subcellular distribution varies depending on the cell type and developmental stage, and therefore the functional features have remained obscure. Although the root plastid proteome is likely to reveal specific functional features, Arabidopsis thaliana root plastid proteome has not been studied to date. RESULTS: In the present study, we separated Arabidopsis root protein fraction enriched with plastids and mitochondria by 2D-PAGE and identified 84 plastid-targeted and 77 mitochondrion-targeted proteins using LC-MS/MS. The most prevalent root plastid protein categories represented amino acid biosynthesis, carbohydrate metabolism and lipid biosynthesis pathways, while the enzymes involved in starch and sucrose metabolism were not detected. Mitochondrion-targeted proteins were classified mainly into the energetics category. CONCLUSIONS: This is the first study presenting gel-based map of Arabidopsis thaliana root plastid and mitochondrial proteome. Our findings suggest that Arabidopsis root plastids have broad biosynthetic capacity, and that they do not play a major role in a long-term storage of carbohydrates. The proteomic map provides a tool for further studies to compare changes in the proteome, e.g. in response to environmental cues, and emphasizes the role of root plastids in nitrogen and sulfur metabolism as well as in amino acid and fatty acid biosynthesis. The results enable taking a first step towards an integrated view of root plastid/mitochondrial proteome and metabolic functions in Arabidopsis thaliana roots.


Asunto(s)
Arabidopsis/genética , Mapeo Cromosómico , Mitocondrias/genética , Proteínas de Plantas/genética , Plastidios/genética , Proteoma/genética , Electroforesis en Gel Bidimensional , Raíces de Plantas/genética , Proteómica
10.
Photosynth Res ; 145(1): 15-30, 2020 Jul.
Artículo en Inglés | MEDLINE | ID: mdl-31975158

RESUMEN

The photosynthetic machinery of plants can acclimate to changes in light conditions by balancing light-harvesting between the two photosystems (PS). This acclimation response is induced by the change in the redox state of the plastoquinone pool, which triggers state transitions through activation of the STN7 kinase and subsequent phosphorylation of light-harvesting complex II (LHCII) proteins. Phosphorylation of LHCII results in its association with PSI (state 2), whereas dephosphorylation restores energy allocation to PSII (state 1). In addition to state transition regulation by phosphorylation, we have recently discovered that plants lacking the chloroplast acetyltransferase NSI are also locked in state 1, even though they possess normal LHCII phosphorylation. This defect may result from decreased lysine acetylation of several chloroplast proteins. Here, we compared the composition of wild type (wt), stn7 and nsi thylakoid protein complexes involved in state transitions separated by Blue Native gel electrophoresis. Protein complex composition and relative protein abundances were determined by LC-MS/MS analyses using iBAQ quantification. We show that despite obvious mechanistic differences leading to defects in state transitions, no major differences were detected in the composition of PSI and LHCII between the mutants. Moreover, both stn7 and nsi plants show retarded growth and decreased PSII capacity under fluctuating light as compared to wt, while the induction of non-photochemical quenching under fluctuating light was much lower in both nsi mutants than in stn7.


Asunto(s)
Aclimatación , Arabidopsis/fisiología , Cloroplastos/metabolismo , Fotosíntesis , Arabidopsis/genética , Cromatografía Liquida , Complejos de Proteína Captadores de Luz/genética , Complejos de Proteína Captadores de Luz/metabolismo , Proteínas Mutantes/metabolismo , Mutación , Oxidación-Reducción , Fosforilación , Complejo de Proteína del Fotosistema I/genética , Complejo de Proteína del Fotosistema I/metabolismo , Complejo de Proteína del Fotosistema II/genética , Complejo de Proteína del Fotosistema II/metabolismo , Plastoquinona/metabolismo , Espectrometría de Masas en Tándem , Tilacoides/metabolismo
11.
Plant Cell ; 28(3): 712-28, 2016 Mar.
Artículo en Inglés | MEDLINE | ID: mdl-26941088

RESUMEN

LIR1 (LIGHT-INDUCED RICE1) encodes a 13-kD, chloroplast-targeted protein containing two nearly identical motifs of unknown function. LIR1 is present in the genomes of vascular plants, mosses, liverworts, and algae, but not in cyanobacteria. Using coimmunoprecipitation assays, pull-down assays, and yeast two-hybrid analyses, we showed that LIR1 interacts with LEAF-TYPE FERREDOXIN-NADP(+) OXIDOREDUCTASE (LFNR), an essential chloroplast enzyme functioning in the last step of photosynthetic linear electron transfer. LIR1 and LFNR formed high molecular weight thylakoid protein complexes with the TIC62 and TROL proteins, previously shown to anchor LFNR to the membrane. We further showed that LIR1 increases the affinity of LFNRs for TIC62 and that the rapid light-triggered degradation of the LIR1 coincides with the release of the LFNR from the thylakoid membrane. Loss of LIR1 resulted in a marked decrease in the accumulation of LFNR-containing thylakoid protein complexes without a concomitant decrease in total LFNR content. In rice (Oryza sativa), photosynthetic capacity of lir1 plants was slightly impaired, whereas no such effect was observed in Arabidopsis thaliana knockout mutants. The consequences of LIR1 deficiency in different species are discussed.


Asunto(s)
Arabidopsis/enzimología , Ferredoxina-NADP Reductasa/metabolismo , Oryza/enzimología , Fotosíntesis , Proteínas de Plantas/metabolismo , Arabidopsis/genética , Arabidopsis/efectos de la radiación , Transporte de Electrón , Ferredoxina-NADP Reductasa/genética , Ferredoxinas/metabolismo , Luz , Complejos Multiproteicos , Mutación , NADP/metabolismo , Oryza/genética , Oryza/efectos de la radiación , Hojas de la Planta/enzimología , Hojas de la Planta/genética , Hojas de la Planta/efectos de la radiación , Proteínas de Plantas/genética , Proteolisis , Especificidad de la Especie , Tilacoides/metabolismo , Técnicas del Sistema de Dos Híbridos
12.
Physiol Plant ; 162(2): 177-190, 2018 Feb.
Artículo en Inglés | MEDLINE | ID: mdl-28833218

RESUMEN

Plastidic ferredoxin-NADP+ oxidoreductases (FNRs; EC:1.18.1.2) together with bacterial type FNRs (FPRs) form the plant-type FNR family. Members of this group contain a two-domain scaffold that forms the basis of an extended superfamily of flavin adenine dinucleotide (FAD) dependent oxidoreductases. In this study, we show that the Arabidopsis thaliana At1g15140 [Ferredoxin-NADP+ oxidoreductase-like (FNRL)] is an FAD-containing NADPH dependent oxidoreductase present in the chloroplast stroma. Determination of the kinetic parameters using the DCPIP NADPH-dependent diaphorase assay revealed that the reaction catalysed by a recombinant FNRL protein followed a saturation Michaelis-Menten profile on the NADPH concentration with kcat = 3.2 ± 0.2 s-1 , KmNADPH = 1.6 ± 0.3 µM and kcat /KmNADPH = 2.0 ± 0.4 µM-1 s-1 . Biochemical assays suggested that FNRL is not likely to interact with Arabidopsis ferredoxin 1, which is supported by the sequence analysis implying that the known Fd-binding residues in plastidic FNRs differ from those of FNRL. In addition, based on structural modelling FNRL has an FAD-binding N-terminal domain built from a six-stranded ß-sheet and one α-helix, and a C-terminal NADP+ -binding α/ß domain with a five-stranded ß-sheet with a pair of α-helices on each side. The FAD-binding site is highly hydrophobic and predicted to bind FAD in a bent conformation typically seen in bacterial FPRs.


Asunto(s)
Proteínas de Arabidopsis/metabolismo , Arabidopsis/enzimología , Proteínas de Cloroplastos/metabolismo , Ferredoxina-NADP Reductasa/metabolismo , Secuencia de Aminoácidos , Arabidopsis/genética , Proteínas de Arabidopsis/química , Proteínas de Arabidopsis/genética , Proteínas Bacterianas/química , Proteínas Bacterianas/genética , Proteínas Bacterianas/metabolismo , Biocatálisis , Proteínas de Cloroplastos/química , Proteínas de Cloroplastos/genética , Ferredoxina-NADP Reductasa/clasificación , Ferredoxina-NADP Reductasa/genética , Flavina-Adenina Dinucleótido/metabolismo , Regulación Enzimológica de la Expresión Génica , Regulación de la Expresión Génica de las Plantas , Cinética , Modelos Moleculares , Filogenia , Dominios Proteicos , Homología de Secuencia de Aminoácido
13.
Photosynth Res ; 134(3): 265-280, 2017 Dec.
Artículo en Inglés | MEDLINE | ID: mdl-28361449

RESUMEN

Ferredoxin-NADP+ reductase (FNR) catalyzes the last step of linear electron transfer in photosynthetic light reactions. The FAD cofactor of FNR accepts two electrons from two independent reduced ferredoxin molecules (Fd) in two sequential steps, first producing neutral semiquinone and then the fully anionic reduced, or hydroquinone, form of the enzyme (FNRhq). FNRhq transfers then both electrons in a single hydride transfer step to NADP+. We are presenting the recent progress in studies focusing on Fd:FNR interaction and subsequent electron transfer processes as well as on interaction of FNR with NADP+/H followed by hydride transfer, both from the structural and functional point of views. We also present the current knowledge about the physiological role(s) of various FNR isoforms present in the chloroplasts of higher plants and the functional impact of subchloroplastic location of FNR. Moreover, open questions and current challenges about the structure, function, and physiology of FNR are discussed.


Asunto(s)
Ferredoxina-NADP Reductasa/química , Ferredoxina-NADP Reductasa/metabolismo , Transporte de Electrón , Ferredoxinas/metabolismo , NADP/metabolismo , Fotosíntesis
14.
Plant Physiol ; 168(3): 768-75, 2015 Jul.
Artículo en Inglés | MEDLINE | ID: mdl-25911530

RESUMEN

Posttranslational modifications of proteins are key effectors of enzyme activity, protein interactions, targeting, and turnover rate, but despite their importance, they are still poorly understood in plants. Although numerous reports have revealed the regulatory role of protein phosphorylation in photosynthesis, various other protein modifications have been identified in chloroplasts only recently. It is known that posttranslational N(α)-acetylation occurs in both nuclear- and plastid-encoded chloroplast proteins, but the physiological significance of this acetylation is not yet understood. Lysine acetylation affects the localization and activity of key metabolic enzymes, and it may work antagonistically or cooperatively with lysine methylation, which also occurs in chloroplasts. In addition, tyrosine nitration may help regulate the repair cycle of photosystem II, while N-glycosylation determines enzyme activity of chloroplastic carbonic anhydrase. This review summarizes the progress in the research field of posttranslational modifications of chloroplast proteins and points out the importance of these modifications in the regulation of chloroplast metabolism.


Asunto(s)
Proteínas de Cloroplastos/metabolismo , Procesamiento Proteico-Postraduccional , Modelos Biológicos
15.
Planta ; 242(3): 733-46, 2015 Sep.
Artículo en Inglés | MEDLINE | ID: mdl-26105652

RESUMEN

MAIN CONCLUSION: The extreme Alb3 C terminus is important for Alb3 stability in a light dependent manner, but is dispensable for LHCP insertion or D1 synthesis. YidC/Oxa1/Alb3 dependent insertion of membrane proteins is evolutionary conserved among bacteria, mitochondria and chloroplasts. Chloroplasts are challenged by the need to coordinate membrane integration of nuclear encoded, post-translationally targeted proteins into the thylakoids as well as of proteins translated on plastid ribosomes. The pathway facilitating post-translational targeting of the light-harvesting chlorophyll a/b binding proteins involves the chloroplast signal recognition particle, cpSRP54 and cpSRP43, as well as its membrane receptor FtsY and the translocase Alb3. Interaction of cpSRP43 with Alb3 is mediated by the positively charged, stromal exposed C terminus of Alb3. In this study, we utilized an Alb3 T-DNA insertion mutant in Arabidopsis thaliana lacking the last 75 amino acids to elucidate the function of this domain (alb3∆C). However, the truncated Alb3 protein (Alb3∆C) proved to be unstable under standard growth conditions, resulting in a reduction of Alb3∆C to 20 % of wild-type levels. In contrast, accumulation of Alb3∆C was comparable to wild type under low light growth conditions. Alb3∆C mutants grown under low light conditions were only slightly paler than wild type, accumulated almost wild-type levels of light harvesting proteins and were not affected in D1 synthesis, therefore showing that the extreme Alb3 C terminus is dispensable for both, co- and post-translational, protein insertion into the thylakoid membrane. However, reduction of Alb3∆C levels as observed under standard growth conditions resulted not only in a severely diminished accumulation of all thylakoid complexes but also in a strong defect in D1 synthesis and membrane insertion.


Asunto(s)
Proteínas de Arabidopsis/química , Proteínas de Arabidopsis/metabolismo , Estabilidad Proteica
16.
New Phytol ; 208(3): 647-55, 2015 Nov.
Artículo en Inglés | MEDLINE | ID: mdl-26174112

RESUMEN

With the tremendous progress of the past decades, molecular plant science is becoming more unified than ever. We now have the exciting opportunity to further connect subdisciplines and understand plants as whole organisms, as will be required to efficiently utilize them in natural and agricultural systems to meet human needs. The subfields of photosynthesis, plant developmental biology and plant stress are used as examples to discuss how plant science can become better integrated. The challenges, strategies and rich opportunities for the integration of the plant sciences are discussed. In recent years, more and more overlap between various subdisciplines has been inadvertently discovered including tradeoffs that may occur in plants engineered for biotechnological applications. Already important, bioinformatics and computational modelling will become even more central to structuring and understanding the ever growing amounts of data. The process of integrating and overlapping fields in plant biology research is advancing, but plant science will benefit from dedicating more effort and urgency to reach across its boundaries.


Asunto(s)
Botánica/tendencias , Fotosíntesis , Desarrollo de la Planta , Plantas/metabolismo , Estrés Fisiológico , Comunicación Celular , Pared Celular/metabolismo , Cloroplastos/metabolismo , Biología Computacional , Expresión Génica , Inmunidad de la Planta , Madera/metabolismo
17.
Plant Physiol ; 166(4): 1764-76, 2014 Dec.
Artículo en Inglés | MEDLINE | ID: mdl-25301888

RESUMEN

Rapid responses of chloroplast metabolism and adjustments to photosynthetic machinery are of utmost importance for plants' survival in a fluctuating environment. These changes may be achieved through posttranslational modifications of proteins, which are known to affect the activity, interactions, and localization of proteins. Recent studies have accumulated evidence about the crucial role of a multitude of modifications, including acetylation, methylation, and glycosylation, in the regulation of chloroplast proteins. Both of the Arabidopsis (Arabidopsis thaliana) leaf-type FERREDOXIN-NADP(+) OXIDOREDUCTASE (FNR) isoforms, the key enzymes linking the light reactions of photosynthesis to carbon assimilation, exist as two distinct forms with different isoelectric points. We show that both AtFNR isoforms contain multiple alternative amino termini and undergo light-responsive addition of an acetyl group to the α-amino group of the amino-terminal amino acid of proteins, which causes the change in isoelectric point. Both isoforms were also found to contain acetylation of a conserved lysine residue near the active site, while no evidence for in vivo phosphorylation or glycosylation was detected. The dynamic, multilayer regulation of AtFNR exemplifies the complex regulatory network systems controlling chloroplast proteins by a range of posttranslational modifications, which continues to emerge as a novel area within photosynthesis research.


Asunto(s)
Arabidopsis/enzimología , Ferredoxina-NADP Reductasa/metabolismo , Procesamiento Proteico-Postraduccional , Acetilación , Secuencia de Aminoácidos , Arabidopsis/genética , Proteínas de Arabidopsis/genética , Proteínas de Arabidopsis/metabolismo , Cloroplastos/enzimología , Ferredoxina-NADP Reductasa/genética , Ferredoxinas/metabolismo , Glicosilación , Isoenzimas , Luz , Modelos Estructurales , Datos de Secuencia Molecular , NADP/metabolismo , Fosforilación , Fotosíntesis , Hojas de la Planta/enzimología , Hojas de la Planta/genética , Alineación de Secuencia
18.
Biochim Biophys Acta ; 1817(1): 247-57, 2012 Jan.
Artículo en Inglés | MEDLINE | ID: mdl-21565160

RESUMEN

The Photosystem (PS) II of cyanobacteria, green algae and higher plants is prone to light-induced inactivation, the D1 protein being the primary target of such damage. As a consequence, the D1 protein, encoded by the psbA gene, is degraded and re-synthesized in a multistep process called PSII repair cycle. In cyanobacteria, a small gene family codes for the various, functionally distinct D1 isoforms. In these organisms, the regulation of the psbA gene expression occurs mainly at the level of transcription, but the expression is fine-tuned by regulation of translation elongation. In plants and green algae, the D1 protein is encoded by a single psbA gene located in the chloroplast genome. In chloroplasts of Chlamydomonas reinhardtii the psbA gene expression is strongly regulated by mRNA processing, and particularly at the level of translation initiation. In chloroplasts of higher plants, translation elongation is the prevalent mechanism for regulation of the psbA gene expression. The pre-existing pool of psbA transcripts forms translation initiation complexes in plant chloroplasts even in darkness, while the D1 synthesis can be completed only in the light. Replacement of damaged D1 protein requires also the assistance by a number of auxiliary proteins, which are encoded by the nuclear genome in green algae and higher plants. Nevertheless, many of these chaperones are conserved between prokaryotes and eukaryotes. Here, we describe the specific features and fundamental differences of the psbA gene expression and the regeneration of the PSII reaction center protein D1 in cyanobacteria, green algae and higher plants. This article is part of a Special Issue entitled Photosystem II.


Asunto(s)
Chlorophyta/genética , Cianobacterias/genética , Complejo de Proteína del Fotosistema II/genética , Plantas/genética , Transcripción Genética , Genes Bacterianos , Genes de Plantas
19.
Plant J ; 70(5): 809-17, 2012 Jun.
Artículo en Inglés | MEDLINE | ID: mdl-22300243

RESUMEN

Arabidopsis thaliana contains two photosynthetically competent chloroplast-targeted ferredoxin-NADP(+) oxidoreductase (FNR) isoforms that are largely redundant in their function. Nevertheless, the FNR isoforms also display distinct molecular phenotypes, as only the FNR1 is able to directly bind to the thylakoid membrane. We report the consequences of depletion of FNR in the F(1) (fnr1 × fnr2) and F(2) (fnr1 fnr2) generation plants of the fnr1 and fnr2 single mutant crossings. The fnr1 × fnr2 plants, with a decreased total content of FNR, showed a small and pale green phenotype, accompanied with a marked downregulation of photosynthetic pigment-protein complexes. Specifically, when compared with the wild type (WT), the quantum yield of photosystem II (PSII) electron transport was lower, non-photochemical quenching (NPQ) was higher and the rate of P700(+) re-reduction was faster in the mutant plants. The slight over-reduction of the plastoquinone pool detected in the mutants resulted in the adjustment of the reactive oxygen species (ROS) scavenging systems, as both the content and de-epoxidation state of xanthophylls, as well as the content of α-tocopherol, were higher in the leaves of the mutant plants when compared with the WT. The fnr1 fnr2 double mutant plants, which had no detectable FNR and possessed an extremely downregulated photosynthetic machinery, survived only when grown heterotrophically in the presence of sucrose. Intriguingly, the fnr1 fnr2 plants were still capable of sustaining the biogenesis of a few malformed chloroplasts.


Asunto(s)
Proteínas de Arabidopsis/metabolismo , Arabidopsis/enzimología , Cloroplastos/enzimología , Ferredoxina-NADP Reductasa/metabolismo , Arabidopsis/anatomía & histología , Arabidopsis/fisiología , Arabidopsis/efectos de la radiación , Proteínas de Arabidopsis/genética , Cloroplastos/fisiología , Cruzamientos Genéticos , Transporte de Electrón , Ferredoxina-NADP Reductasa/genética , Isoenzimas/genética , Isoenzimas/metabolismo , Luz , Microscopía Electrónica de Transmisión , Oxidación-Reducción , Estrés Oxidativo , Fenotipo , Fotosíntesis , Complejo de Proteína del Fotosistema II/genética , Complejo de Proteína del Fotosistema II/metabolismo , Hojas de la Planta/metabolismo , Hojas de la Planta/efectos de la radiación , Hojas de la Planta/ultraestructura , Especies Reactivas de Oxígeno/metabolismo , Sacarosa , Tilacoides/genética , Tilacoides/metabolismo , Xantófilas/genética , Xantófilas/metabolismo
20.
Plant Cell Environ ; 36(6): 1071-84, 2013 Jun.
Artículo en Inglés | MEDLINE | ID: mdl-23190083

RESUMEN

Ferredoxin (Fd) is a small [2Fe-2S] cluster-containing protein found in all organisms performing oxygenic photosynthesis. Fd is the first soluble acceptor of electrons on the stromal side of the chloroplast electron transport chain, and as such is pivotal to determining the distribution of these electrons to different metabolic reactions. In chloroplasts, the principle sink for electrons is in the production of NADPH, which is mostly consumed during the assimilation of CO2 . In addition to this primary function in photosynthesis, Fds are also involved in a number of other essential metabolic reactions, including biosynthesis of chlorophyll, phytochrome and fatty acids, several steps in the assimilation of sulphur and nitrogen, as well as redox signalling and maintenance of redox balance via the thioredoxin system and Halliwell-Asada cycle. This makes Fds crucial determinants of the electron transfer between the thylakoid membrane and a variety of soluble enzymes dependent on these electrons. In this article, we will first describe the current knowledge on the structure and function of the various Fd isoforms present in chloroplasts of higher plants and then discuss the processes involved in oxidation of Fd, introducing the corresponding enzymes and discussing what is known about their relative interaction with Fd.


Asunto(s)
Cloroplastos/metabolismo , Ferredoxinas/metabolismo , Fotosíntesis , Plantas/metabolismo , Cloroplastos/fisiología , Transporte de Electrón/fisiología , Ferredoxinas/fisiología , Fotosíntesis/fisiología
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