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  1 / 136 MEDLINE  
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[PMID]:28530801
[Au] Autor:Iwata T; Nozaki D; Yamamoto A; Koyama T; Nishina Y; Shiga K; Tokutomi S; Unno M; Kandori H
[Ad] Endereço:Department of Life Science and Applied Chemistry, Nagoya Institute of Technology , Showa-ku, Nagoya 466-8555, Japan.
[Ti] Título:Hydrogen Bonding Environment of the N3-H Group of Flavin Mononucleotide in the Light Oxygen Voltage Domains of Phototropins.
[So] Source:Biochemistry;56(24):3099-3108, 2017 Jun 20.
[Is] ISSN:1520-4995
[Cp] País de publicação:United States
[La] Idioma:eng
[Ab] Resumo:The light oxygen voltage (LOV) domain is a flavin-binding blue-light receptor domain, originally found in a plant photoreceptor phototropin (phot). Recently, LOV domains have been used in optogenetics as the photosensory domain of fusion proteins. Therefore, it is important to understand how LOV domains exhibit light-induced structural changes for the kinase domain regulation, which enables the design of LOV-containing optogenetics tools with higher photoactivation efficiency. In this study, the hydrogen bonding environment of the N3-H group of flavin mononucleotide (FMN) of the LOV2 domain from Adiantum neochrome (neo) 1 was investigated by low-temperature Fourier transform infrared spectroscopy. Using specifically N-labeled FMN, [1,3- N ]FMN, the N3-H stretch was identified at 2831 cm for the unphotolyzed state at 150 K, indicating that the N3-H group forms a fairly strong hydrogen bond. The N3-H stretch showed temperature dependence, with a shift to lower frequencies at ≤200 K and to higher frequencies at ≥250 K from the unphotolyzed to the intermediate states. Similar trends were observed in the LOV2 domains from Arabidopsis phot1 and phot2. By contrast, the N3-H stretch of the Q1029L mutant of neo1-LOV2 and neo1-LOV1 was not temperature dependent in the intermediate state. These results seemed correlated with our previous finding that the LOV2 domains show the structural changes in the ß-sheet region and/or the adjacent Jα helix of LOV2 domain, but that such structural changes do not take place in the Q1029L mutant or neo1-LOV1 domain. The environment around the N3-H group was also investigated.
[Mh] Termos MeSH primário: Proteínas de Arabidopsis/metabolismo
Proteínas de Ligação a DNA/metabolismo
Mononucleotídeo de Flavina/química
Mononucleotídeo de Flavina/metabolismo
Fototropinas/química
Fototropinas/metabolismo
[Mh] Termos MeSH secundário: Proteínas de Arabidopsis/química
Proteínas de Ligação a DNA/química
Ligações de Hidrogênio
[Pt] Tipo de publicação:JOURNAL ARTICLE
[Nm] Nome de substância:
0 (Arabidopsis Proteins); 0 (DNA-Binding Proteins); 0 (LOV2 protein, Arabidopsis); 0 (Phototropins); 7N464URE7E (Flavin Mononucleotide)
[Em] Mês de entrada:1709
[Cu] Atualização por classe:170926
[Lr] Data última revisão:
170926
[Sb] Subgrupo de revista:IM
[Da] Data de entrada para processamento:170523
[St] Status:MEDLINE
[do] DOI:10.1021/acs.biochem.7b00057


  2 / 136 MEDLINE  
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[PMID]:28412349
[Au] Autor:Sun W; Zhang W; Zhang C; Mao M; Zhao Y; Chen X; Yang Y
[Ad] Endereço:Synthetic Biology and Biotechnology Laboratory, State Key Laboratory of Bioreactor Engineering, School of Pharmacy, East China University of Science and Technology, 130 Mei Long Road, Shanghai 200237, China.
[Ti] Título:Light-induced protein degradation in human-derived cells.
[So] Source:Biochem Biophys Res Commun;487(2):241-246, 2017 May 27.
[Is] ISSN:1090-2104
[Cp] País de publicação:United States
[La] Idioma:eng
[Ab] Resumo:Controlling protein degradation can be a valuable tool for posttranslational regulation of protein abundance to study complex biological systems. In the present study, we designed a light-switchable degron consisting of a light oxygen voltage (LOV) domain of Avena sativa phototropin 1 (AsLOV2) and a C-terminal degron. Our results showed that the light-switchable degron could be used for rapid and specific induction of protein degradation in HEK293 cells by light in a proteasome-dependent manner. Further studies showed that the light-switchable degron could also be utilized to mediate the degradation of secreted Gaussia princeps luciferase (GLuc), demonstrating the adaptability of the light-switchable degron in different types of protein. We suggest that the light-switchable degron offers a robust tool to control protein levels and may serves as a new and significant method for gene- and cell-based therapies.
[Mh] Termos MeSH primário: Avena/metabolismo
Luz
Medições Luminescentes/métodos
Fototropinas/metabolismo
Proteólise/efeitos da radiação
[Mh] Termos MeSH secundário: Relação Dose-Resposta à Radiação
Genes Reporter/fisiologia
Células HEK293
Seres Humanos
Dose de Radiação
[Pt] Tipo de publicação:JOURNAL ARTICLE
[Nm] Nome de substância:
0 (Phototropins)
[Em] Mês de entrada:1706
[Cu] Atualização por classe:170612
[Lr] Data última revisão:
170612
[Sb] Subgrupo de revista:IM
[Da] Data de entrada para processamento:170417
[St] Status:MEDLINE


  3 / 136 MEDLINE  
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[PMID]:27631339
[Au] Autor:Deng Z; Wang ZY; Kutschera U
[Ad] Endereço:State Key Laboratory Breeding Base for Zhejiang Sustainable Pest and Disease Control, Institute of Virology and Biotechnology, Zhejiang Academy of Agricultural Sciences, Hangzhou, 310021, China.
[Ti] Título:Seedling development in maize cv. B73 and blue light-mediated proteomic changes in the tip vs. stem of the coleoptile.
[So] Source:Protoplasma;254(3):1317-1322, 2017 May.
[Is] ISSN:1615-6102
[Cp] País de publicação:Austria
[La] Idioma:eng
[Ab] Resumo:In 2009, the draft genome of the reference inbred line of maize (Zea mays L. spp. mays cv. B73) was published so that, using this specific corn variety, molecular analyses of physiological processes became possible. However, the morphology and developmental patterns of B73 maize, compared with that of the more frequently used hybrid varieties, have not yet been analyzed. Here, we describe organ development in seedlings of B73 maize and in those of six other hybrid cultivars, and document significant morphological as well as quantitative differences between these varieties of Z. mays. In a second set of experiments, we used etiolated seedlings of B73 maize to analyze the effect of blue light (BL) on the patterns of proteins in the tip vs. growing region of this sheath-like organ. By using two-dimensional difference gel electrophoresis (2D DIGE), coupled with tandem mass spectrometry, we detected, in the microsomal fraction of maize coleoptile tips, rapid changes in the abundance of protein spots of maize phototropin 1 and several metabolic enzymes. In the sub-apical (growing) region of the coleoptile, proteomic changes were less pronounced. These results suggest that the tip of the coleoptile of B73 maize may serve as a unique model system for dissecting BL responses in a light-sensitive plant organ of known function.
[Mh] Termos MeSH primário: Cotilédone/metabolismo
Luz
Proteínas de Plantas/metabolismo
Caules de Planta/metabolismo
Plântulas/crescimento & desenvolvimento
Zea mays/embriologia
[Mh] Termos MeSH secundário: Quimera/crescimento & desenvolvimento
Glucosiltransferases/metabolismo
Lipoxigenases/metabolismo
Fototropinas/metabolismo
Proteoma/metabolismo
Proteômica
Plântulas/embriologia
Zea mays/metabolismo
[Pt] Tipo de publicação:JOURNAL ARTICLE
[Nm] Nome de substância:
0 (Phototropins); 0 (Plant Proteins); 0 (Proteome); EC 1.13.11.- (Lipoxygenases); EC 2.4.1.- (Glucosyltransferases); EC 2.4.1.13 (sucrose synthase)
[Em] Mês de entrada:1705
[Cu] Atualização por classe:171012
[Lr] Data última revisão:
171012
[Sb] Subgrupo de revista:IM
[Da] Data de entrada para processamento:160916
[St] Status:MEDLINE
[do] DOI:10.1007/s00709-016-1023-6


  4 / 136 MEDLINE  
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[PMID]:27435584
[Au] Autor:Souslova EA; Mironova KE; Deyev SM
[Ad] Endereço:Shemyakin & Ovchinnikov Institute of Bioorganic Chemistry of Russian Academy of Sciences (IBCH RAS), Miklukho-Maklaya str. 16/10, Moscow, 117997, Russia.
[Ti] Título:Applications of genetically encoded photosensitizer miniSOG: from correlative light electron microscopy to immunophotosensitizing.
[So] Source:J Biophotonics;10(3):338-352, 2017 Mar.
[Is] ISSN:1864-0648
[Cp] País de publicação:Germany
[La] Idioma:eng
[Ab] Resumo:Genetically encoded photosensitizers (PSs), e.g. ROS generating proteins, correspond to a novel class of PSs that are highly desirable for biological and medical applications since they can be used in combination with a variety of genetic engineering manipulations allowing for precise spatio-temporal control of ROS production within living cells and organisms. In contrast to the commonly used chemical PSs, they can be modified using genetic engineering approaches and targeted to particular cellular compartments and cell types. Mini Singlet Oxygen Generator (miniSOG), a small flavoprotein capable of singlet oxygen production upon blue light irradiation, was initially reported as a high contrast probe for correlative light electron microscopy (CLEM) without the need of exogenous ligands, probes or destructive permeabilizing detergents. Further miniSOG was successfully applied for chromophore-assisted light inactivation (CALI) of proteins, as well as for photo-induced cell ablation in tissue cultures and in Caenorhabditis elegans. Finally, a novel approach of immunophotosensitizing has been developed, exploiting the specificity of mini-antibodies or selective scaffold proteins and photo-induced cytotoxicity of miniSOG, which is particularly promising for selective non-invasive photodynamic therapy of cancer (PDT) due to the spatial selectivity and locality of destructive action compared to other methods of oncotherapy.
[Mh] Termos MeSH primário: Meios de Contraste
Flavoproteínas
Fármacos Fotossensibilizantes
Fototropinas
[Mh] Termos MeSH secundário: Animais
Antineoplásicos/farmacologia
Flavoproteínas/química
Flavoproteínas/genética
Flavoproteínas/farmacologia
Seres Humanos
Luz
Microscopia Eletrônica
Fármacos Fotossensibilizantes/farmacologia
Fototropinas/química
Fototropinas/genética
Fototropinas/farmacologia
Espécies Reativas de Oxigênio/metabolismo
Oxigênio Singlete/metabolismo
[Pt] Tipo de publicação:JOURNAL ARTICLE; REVIEW
[Nm] Nome de substância:
0 (Antineoplastic Agents); 0 (Contrast Media); 0 (Flavoproteins); 0 (Photosensitizing Agents); 0 (Phototropins); 0 (Reactive Oxygen Species); 17778-80-2 (Singlet Oxygen)
[Em] Mês de entrada:1710
[Cu] Atualização por classe:171010
[Lr] Data última revisão:
171010
[Sb] Subgrupo de revista:IM
[Da] Data de entrada para processamento:160721
[St] Status:MEDLINE
[do] DOI:10.1002/jbio.201600120


  5 / 136 MEDLINE  
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[PMID]:27717456
[Au] Autor:Vanhaelewyn L; Schumacher P; Poelman D; Fankhauser C; Van Der Straeten D; Vandenbussche F
[Ad] Endereço:Laboratory of Functional Plant Biology, Department of Physiology, Faculty of Sciences, Ghent University, KL Ledeganckstraat 35, B-9000 Gent, Belgium.
[Ti] Título:REPRESSOR OF ULTRAVIOLET-B PHOTOMORPHOGENESIS function allows efficient phototropin mediated ultraviolet-B phototropism in etiolated seedlings.
[So] Source:Plant Sci;252:215-221, 2016 Nov.
[Is] ISSN:1873-2259
[Cp] País de publicação:Ireland
[La] Idioma:eng
[Ab] Resumo:Ultraviolet B (UV-B) light is a part of the solar radiation which has significant effects on plant morphology, even at low doses. In Arabidopsis, many of these morphological changes have been attributed to a specific UV-B receptor, UV resistance locus 8 (UVR8). Recent findings showed that next to phototropin regulated phototropism, UVR8 mediated signaling is able of inducing directional bending towards UV-B light in etiolated seedlings of Arabidopsis, in a phototropin independent manner. In this study, kinetic analysis of phototropic bending was used to evaluate the relative contribution of each of these pathways in UV-B mediated phototropism. Diminishing UV-B light intensity favors the importance of phototropins. Molecular and genetic analyses suggest that UV-B is capable of inducing phototropin signaling relying on phototropin kinase activity and regulation of NPH3. Moreover, enhanced UVR8 responses in the UV-B hypersensitive rup1rup2 mutants interferes with the fast phototropin mediated phototropism. Together the data suggest that phototropins are the most important receptors for UV-B induced phototropism in etiolated seedlings, and a RUP mediated negative feedback pathway prevents UVR8 signaling to interfere with the phototropin dependent response.
[Mh] Termos MeSH primário: Arabidopsis/efeitos da radiação
Fototropinas/fisiologia
Fototropismo
Raios Ultravioleta
[Mh] Termos MeSH secundário: Arabidopsis/crescimento & desenvolvimento
Arabidopsis/metabolismo
Proteínas de Arabidopsis/genética
Proteínas de Arabidopsis/metabolismo
Proteínas de Arabidopsis/fisiologia
Proteínas Cromossômicas não Histona/genética
Proteínas Cromossômicas não Histona/metabolismo
Proteínas Cromossômicas não Histona/fisiologia
Cinética
Transdução de Sinal Luminoso
Fototropinas/genética
Fototropinas/metabolismo
Plântulas/crescimento & desenvolvimento
Plântulas/metabolismo
Plântulas/efeitos da radiação
[Pt] Tipo de publicação:JOURNAL ARTICLE
[Nm] Nome de substância:
0 (Arabidopsis Proteins); 0 (Chromosomal Proteins, Non-Histone); 0 (Phototropins); 0 (Uvr8 protein, Arabidopsis)
[Em] Mês de entrada:1704
[Cu] Atualização por classe:170417
[Lr] Data última revisão:
170417
[Sb] Subgrupo de revista:IM
[Da] Data de entrada para processamento:161009
[St] Status:MEDLINE


  6 / 136 MEDLINE  
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[PMID]:27626383
[Au] Autor:Petroutsos D; Tokutsu R; Maruyama S; Flori S; Greiner A; Magneschi L; Cusant L; Kottke T; Mittag M; Hegemann P; Finazzi G; Minagawa J
[Ad] Endereço:Laboratoire de Physiologie Cellulaire et Végétale, UMR 5168, Centre National de la Recherche Scientifique, Commissariat à l'Energie Atomique et aux Energies Alternatives, Université Grenoble Alpes, Institut National Recherche Agronomique, Institut de Biosciences et Biotechnologies de Grenoble, (BIG)
[Ti] Título:A blue-light photoreceptor mediates the feedback regulation of photosynthesis.
[So] Source:Nature;537(7621):563-566, 2016 09 22.
[Is] ISSN:1476-4687
[Cp] País de publicação:England
[La] Idioma:eng
[Ab] Resumo:In plants and algae, light serves both as the energy source for photosynthesis and a biological signal that triggers cellular responses via specific sensory photoreceptors. Red light is perceived by bilin-containing phytochromes and blue light by the flavin-containing cryptochromes and/or phototropins (PHOTs), the latter containing two photosensory light, oxygen, or voltage (LOV) domains. Photoperception spans several orders of light intensity, ranging from far below the threshold for photosynthesis to values beyond the capacity of photosynthetic CO assimilation. Excess light may cause oxidative damage and cell death, processes prevented by enhanced thermal dissipation via high-energy quenching (qE), a key photoprotective response. Here we show the existence of a molecular link between photoreception, photosynthesis, and photoprotection in the green alga Chlamydomonas reinhardtii. We show that PHOT controls qE by inducing the expression of the qE effector protein LHCSR3 (light-harvesting complex stress-related protein 3) in high light intensities. This control requires blue-light perception by LOV domains on PHOT, LHCSR3 induction through PHOT kinase, and light dissipation in photosystem II via LHCSR3. Mutants deficient in the PHOT gene display severely reduced fitness under excessive light conditions, indicating that the sensing, utilization, and dissipation of light is a concerted process that plays a vital role in microalgal acclimation to environments of variable light intensities.
[Mh] Termos MeSH primário: Chlamydomonas reinhardtii/metabolismo
Chlamydomonas reinhardtii/efeitos da radiação
Retroalimentação Fisiológica/efeitos da radiação
Transdução de Sinal Luminoso/efeitos da radiação
Luz
Fotossíntese/efeitos da radiação
Fototropinas/metabolismo
[Mh] Termos MeSH secundário: Aclimatação/efeitos da radiação
Sobrevivência Celular/efeitos da radiação
Chlamydomonas reinhardtii/genética
Cor
Complexos de Proteínas Captadores de Luz/biossíntese
Complexos de Proteínas Captadores de Luz/metabolismo
Complexo de Proteína do Fotossistema II/metabolismo
Fototropinas/química
Fototropinas/genética
Proteínas Quinases/química
Proteínas Quinases/metabolismo
[Pt] Tipo de publicação:JOURNAL ARTICLE; RESEARCH SUPPORT, NON-U.S. GOV'T
[Nm] Nome de substância:
0 (Light-Harvesting Protein Complexes); 0 (Photosystem II Protein Complex); 0 (Phototropins); EC 2.7.- (Protein Kinases)
[Em] Mês de entrada:1702
[Cu] Atualização por classe:170815
[Lr] Data última revisão:
170815
[Sb] Subgrupo de revista:IM
[Da] Data de entrada para processamento:160915
[St] Status:MEDLINE
[do] DOI:10.1038/nature19358


  7 / 136 MEDLINE  
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[PMID]:27586333
[Au] Autor:Zimmerman SP; Kuhlman B; Yumerefendi H
[Ad] Endereço:University of North Carolina Chapel Hill, Chapel Hill, NC, United States.
[Ti] Título:Engineering and Application of LOV2-Based Photoswitches.
[So] Source:Methods Enzymol;580:169-90, 2016.
[Is] ISSN:1557-7988
[Cp] País de publicação:United States
[La] Idioma:eng
[Ab] Resumo:Cellular optogenetic switches, a novel class of biological tools, have improved our understanding of biological phenomena that were previously intractable. While the design and engineering of these proteins has historically varied, they are all based on borrowed elements from plant and bacterial photoreceptors. In general terms, each of the optogenetic switches designed to date exploits the endogenous light-induced change in photoreceptor conformation while repurposing its effect to target a different biological phenomenon. We focus on the well-characterized light-oxygen-voltage 2 (LOV2) domain from Avena sativa phototropin 1 as our cornerstone for design. While the function of the LOV2 domain in the context of the phototropin protein is not fully elucidated, its thorough biophysical characterization as an isolated domain has created a strong foundation for engineering of photoswitches. In this chapter, we examine the biophysical characteristics of the LOV2 domain that may be exploited to produce an optogenetic switch and summarize previous design efforts to provide guidelines for an effective design. Furthermore, we provide protocols for assays including fluorescence polarization, phage display, and microscopy that are optimized for validating, improving, and using newly designed photoswitches.
[Mh] Termos MeSH primário: Flavoproteínas/química
Luz
Fototropinas/química
Engenharia de Proteínas/métodos
[Mh] Termos MeSH secundário: Avena/enzimologia
Modelos Moleculares
Estrutura Terciária de Proteína/efeitos da radiação
[Pt] Tipo de publicação:JOURNAL ARTICLE
[Nm] Nome de substância:
0 (Flavoproteins); 0 (Phototropins)
[Em] Mês de entrada:1708
[Cu] Atualização por classe:170807
[Lr] Data última revisão:
170807
[Sb] Subgrupo de revista:IM
[Da] Data de entrada para processamento:160903
[St] Status:MEDLINE


  8 / 136 MEDLINE  
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[PMID]:27406783
[Au] Autor:Sztatelman O; Labuz J; Hermanowicz P; Banas AK; Bazant A; Zglobicki P; Aggarwal C; Nadzieja M; Krzeszowiec W; Strzalka W; Gabrys H
[Ad] Endereço:Department of Plant Biotechnology, Faculty of Biochemistry, Biophysics and Biotechnology, Jagiellonian University, Gronostajowa 7, 30-387 Krakow, Poland Institute of Biochemistry and Biophysics, Polish Academy of Sciences, Pawinskiego 5a, 02-106 Warsaw, Poland olga.sztatelman@ibb.waw.pl.
[Ti] Título:Fine tuning chloroplast movements through physical interactions between phototropins.
[So] Source:J Exp Bot;67(17):4963-78, 2016 Sep.
[Is] ISSN:1460-2431
[Cp] País de publicação:England
[La] Idioma:eng
[Ab] Resumo:Phototropins are plant photoreceptors which regulate numerous responses to blue light, including chloroplast relocation. Weak blue light induces chloroplast accumulation, whereas strong light leads to an avoidance response. Two Arabidopsis phototropins are characterized by different light sensitivities. Under continuous light, both can elicit chloroplast accumulation, but the avoidance response is controlled solely by phot2. As well as continuous light, brief light pulses also induce chloroplast displacements. Pulses of 0.1s and 0.2s of fluence rate saturating the avoidance response lead to transient chloroplast accumulation. Longer pulses (up to 20s) trigger a biphasic response, namely transient avoidance followed by transient accumulation. This work presents a detailed study of transient chloroplast responses in Arabidopsis. Phototropin mutants display altered chloroplast movements as compared with the wild type: phot1 is characterized by weaker responses, while phot2 exhibits enhanced chloroplast accumulation, especially after 0.1s and 0.2s pulses. To determine the cause of these differences, the abundance and phosphorylation levels of both phototropins, as well as the interactions between phototropin molecules are examined. The formation of phototropin homo- and heterocomplexes is the most plausible explanation of the observed phenomena. The physiological consequences of this interplay are discussed, suggesting the universal character of this mechanism that fine-tunes plant reactions to blue light. Additionally, responses in mutants of different protein phosphatase 2A subunits are examined to assess the role of protein phosphorylation in signaling of chloroplast movements.
[Mh] Termos MeSH primário: Cloroplastos/fisiologia
Fototropinas/fisiologia
[Mh] Termos MeSH secundário: Arabidopsis/metabolismo
Arabidopsis/fisiologia
Arabidopsis/efeitos da radiação
Proteínas de Arabidopsis/metabolismo
Proteínas de Arabidopsis/fisiologia
Cloroplastos/metabolismo
Cloroplastos/efeitos da radiação
Luz
Fototropinas/metabolismo
Técnicas do Sistema de Duplo-Híbrido
[Pt] Tipo de publicação:JOURNAL ARTICLE
[Nm] Nome de substância:
0 (Arabidopsis Proteins); 0 (Phototropins)
[Em] Mês de entrada:1711
[Cu] Atualização por classe:171107
[Lr] Data última revisão:
171107
[Sb] Subgrupo de revista:IM
[Da] Data de entrada para processamento:160714
[St] Status:MEDLINE
[do] DOI:10.1093/jxb/erw265


  9 / 136 MEDLINE  
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[PMID]:27310016
[Au] Autor:Suetsugu N; Higa T; Gotoh E; Wada M
[Ad] Endereço:Department of Biology, Faculty of Sciences, Kyushu University, Fukuoka, Japan.
[Ti] Título:Light-Induced Movements of Chloroplasts and Nuclei Are Regulated in Both Cp-Actin-Filament-Dependent and -Independent Manners in Arabidopsis thaliana.
[So] Source:PLoS One;11(6):e0157429, 2016.
[Is] ISSN:1932-6203
[Cp] País de publicação:United States
[La] Idioma:eng
[Ab] Resumo:Light-induced chloroplast movement and attachment to the plasma membrane are dependent on actin filaments. In Arabidopsis thaliana, the short actin filaments on the chloroplast envelope, cp-actin filaments, are essential for chloroplast movement and positioning. Furthermore, cp-actin-filament-mediated chloroplast movement is necessary for the strong-light-induced nuclear avoidance response. The proteins CHLOROPLAST UNUSUAL POSITIONING 1 (CHUP1), KINESIN-LIKE PROTEIN FOR ACTIN-BASED CHLOROPLAST MOVEMENT 1 (KAC1) and KAC2 are required for the generation and/or maintenance of cp-actin filaments in Arabidopsis. In land plants, CHUP1 and KAC family proteins play pivotal roles in the proper movement of chloroplasts and their attachment to the plasma membrane. Here, we report similar but distinct phenotypes in chloroplast and nuclear photorelocation movements between chup1 and kac1kac2 mutants. Measurement of chloroplast photorelocation movement indicated that kac1kac2, but not chup1, exhibited a clear strong-light-induced increase in leaf transmittance changes. The chloroplast movement in kac1kac2 depended on phototropin 2, CHUP1 and two other regulators for cp-actin filaments, PLASTID MOVEMENT IMPAIRED 1 and THRUMIN 1. Furthermore, kac1kac2 retained a weak but significant nuclear avoidance response although chup1 displayed a severe defect in the nuclear avoidance response. The kac1kac2chup1 triple mutant was completely defective in both chloroplast and nuclear avoidance responses. These results indicate that CHUP1 and the KACs function somewhat independently, but interdependently mediate both chloroplast and nuclear photorelocation movements.
[Mh] Termos MeSH primário: Proteínas de Arabidopsis/genética
Arabidopsis/efeitos da radiação
Núcleo Celular/efeitos da radiação
Proteínas de Cloroplastos/genética
Cloroplastos/efeitos da radiação
Regulação da Expressão Gênica de Plantas
Cinesina/genética
Proteínas dos Microfilamentos/genética
[Mh] Termos MeSH secundário: Citoesqueleto de Actina/metabolismo
Citoesqueleto de Actina/efeitos da radiação
Citoesqueleto de Actina/ultraestrutura
Arabidopsis/genética
Arabidopsis/metabolismo
Arabidopsis/ultraestrutura
Proteínas de Arabidopsis/metabolismo
Membrana Celular/metabolismo
Membrana Celular/efeitos da radiação
Membrana Celular/ultraestrutura
Núcleo Celular/metabolismo
Núcleo Celular/ultraestrutura
Proteínas de Cloroplastos/metabolismo
Cloroplastos/metabolismo
Cloroplastos/ultraestrutura
Peptídeos e Proteínas de Sinalização Intracelular/genética
Peptídeos e Proteínas de Sinalização Intracelular/metabolismo
Cinesina/metabolismo
Luz
Proteínas dos Microfilamentos/metabolismo
Movimento
Fototropinas/genética
Fototropinas/metabolismo
Células Vegetais/metabolismo
Células Vegetais/efeitos da radiação
Células Vegetais/ultraestrutura
Folhas de Planta/genética
Folhas de Planta/metabolismo
Folhas de Planta/efeitos da radiação
Folhas de Planta/ultraestrutura
[Pt] Tipo de publicação:JOURNAL ARTICLE
[Nm] Nome de substância:
0 (Arabidopsis Proteins); 0 (Chloroplast Proteins); 0 (Intracellular Signaling Peptides and Proteins); 0 (Microfilament Proteins); 0 (Phototropins); 0 (Pmi1 protein, Arabidopsis); 0 (THRUMIN1 protein, Arabidopsis); 0 (chloroplast unusual positioning1 protein, Arabidopsis); EC 3.6.1.- (KAC1 protein, Arabidopsis); EC 3.6.1.- (KAC2 protein, Arabidopsis); EC 3.6.4.4 (Kinesin)
[Em] Mês de entrada:1707
[Cu] Atualização por classe:170713
[Lr] Data última revisão:
170713
[Sb] Subgrupo de revista:IM
[Da] Data de entrada para processamento:160617
[St] Status:MEDLINE
[do] DOI:10.1371/journal.pone.0157429


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[PMID]:27295881
[Au] Autor:Zou X; Xiao R; Guo X; Qu J; Lu Z; Hong T
[Ti] Título:[Preparation of Recombinant Human Adenoviruses Labeled with miniSOG].
[So] Source:Bing Du Xue Bao;32(1):32-8, 2016 Jan.
[Is] ISSN:1000-8721
[Cp] País de publicação:China
[La] Idioma:chi
[Ab] Resumo:We wished to study the intracellular transport of adenoviruses. We constructed a novel recombinant adenovirus in which the structural protein IX was labeled with a mini-singlet oxygen generator (miniSOG). The miniSOG gene was synthesized by overlapping extension polymerase chain reaction (PCR), cloned to the pcDNA3 vector, and expressed in 293 cells. Activation of miniSOG generated sufficient numbers of singlet oxygen molecules to catalyze polymerization of diaminobenzidine into an osmiophilic reaction product resolvable by transmission electron microscopy (TEM). To construct miniSOG-labelled recombinant adenoviruses, the miniSOG gene was subcloned downstream of the IX gene in a pShuttle plasmid. Adenoviral plasmid pAd5-IXSOG was generated by homologous recombination of the modified shuttle plasmid (pShuttle-IXSOG) with the backbone plasmid (pAdeasy-1) in the BJ5183 strain of Eschericia coli. Adenovirus HAdV-5-IXSOG was rescued by transfection of 293 cells with the linearized pAd5-IXSOG. After propagation, virions were purified using the CsC1 ultracentrifugation method. Finally, HAdV-5-IXSOG in 2.0 mL with a particle titer of 6 x 1011 vp/mL was obtained. Morphology of HAdV-5-IXSOG was verified by TEM. Fusion of IX with the miniSOG gene was confirmed by PCR. In conclusion, miniSOG-labeled recombinant adenoviruses were constructed, which could be valuable tools for virus tracking by TEM.
[Mh] Termos MeSH primário: Adenovírus Humanos/química
Proteínas de Arabidopsis/química
Flavoproteínas/química
[Mh] Termos MeSH secundário: Adenovírus Humanos/genética
Adenovírus Humanos/metabolismo
Proteínas de Arabidopsis/genética
Proteínas de Arabidopsis/metabolismo
Flavoproteínas/genética
Flavoproteínas/metabolismo
Seres Humanos
Fototropinas/química
Fototropinas/genética
Fototropinas/metabolismo
Oxigênio Singlete/química
Coloração e Rotulagem
Transfecção
[Pt] Tipo de publicação:ENGLISH ABSTRACT; JOURNAL ARTICLE; RESEARCH SUPPORT, NON-U.S. GOV'T
[Nm] Nome de substância:
0 (Arabidopsis Proteins); 0 (Flavoproteins); 0 (Phototropins); 17778-80-2 (Singlet Oxygen)
[Em] Mês de entrada:1606
[Cu] Atualização por classe:161021
[Lr] Data última revisão:
161021
[Sb] Subgrupo de revista:IM
[Da] Data de entrada para processamento:160615
[St] Status:MEDLINE



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