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Biochimica et biophysica acta, General subjects 21건

  1. [해외논문]   Role of potassium channels in chlorogenic acid-induced apoptotic volume decrease and cell cycle arrest in Candida albicans   SCI SCIE

    Yun, JiEun (Corresponding author.) , Lee, Dong Gun
    Biochimica et biophysica acta, General subjects v.1861 no.3 ,pp. 585 - 592 , 2017 , 0304-4165 ,

    초록

    Abstract Background Chlorogenic acid (CRA) is an abundant phenolic compound in the human diet. CRA has a potent antifungal effect, inducing cell death in Candida albicans . However, there are no further studies to investigate the antifungal mechanism of CRA, associated with ion channels. Methods To evaluate the inhibitory effects on CRA-induced cell death, C. albicans cells were pretreated with potassium and chloride channel blockers, separately. Flow cytometry was carried out to detect several hallmarks of apoptosis, such as cell cycle arrest, caspase activation, and DNA fragmentation, after staining of the cells with SYTOX green, FITC-VAD-FMK, and TUNEL. Results CRA caused excessive potassium efflux, and an apoptotic volume decrease (AVD) was observed. This change, in turn, induced cytosolic calcium uptake and cell cycle arrest in C. albicans . Moreover, CRA induced caspase activation and DNA fragmentation, which are considered apoptotic markers. In contrast, the potassium efflux and proapoptotic changes were inhibited when potassium channels were blocked, whereas there was no inhibitory effect when chloride channels were blocked. Conclusions CRA induces potassium efflux, leading to AVD and G2/M cell cycle arrest in C. albicans . Therefore, potassium efflux via potassium channels regulates the CRA-induced apoptosis, stimulating several apoptotic processes. General significance This study improves the understanding of the antifungal mechanism of CRA and its association with ion homeostasis, thereby pointing to a role of potassium channels in CRA-induced apoptosis. Highlights Chlorogenic acid (CRA) induces apoptosis in Candida albicans . CRA induces potassium efflux leading to apoptotic volume decrease. Block of the potassium channels inhibits the apoptotic effect of CRA. Potassium channels play a key role in regulation of the CRA-induced apoptosis.

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  2. [해외논문]   Putative DNA modification methylase DR_C0020 of Deinococcus radiodurans is an atypical SAM dependent C-5 cytosine DNA methylase   SCI SCIE

    Patil, Nayana A. (Molecular Biology Division, Bhabha Atomic Research Centre, Mumbai 400 085, India ) , Basu, Bhakti (Molecular Biology Division, Bhabha Atomic Research Centre, Mumbai 400 085, India ) , Deobagkar, Deepti D. (Department of Zoology, Centre for Advanced Studies, Savitribai Phule Pune University, Pune 411007, India ) , Apte, Shree K. (Molecular Biology Division, Bhabha Atomic Research Centre, Mumbai 400 085, India ) , Deobagkar, Dileep N. (Department of Zoology, Centre for Advanced Studies, Savitribai Phule Pune University, Pune 411007, India)
    Biochimica et biophysica acta, General subjects v.1861 no.3 ,pp. 593 - 602 , 2017 , 0304-4165 ,

    초록

    Abstract Background Control of cellular processes by epigenetic modification of cytosine in DNA is widespread among living organisms, but, is hitherto unknown in the extremely radioresistant microbe D. radiodurans . Methods C-5 methyl cytosines (m 5 C) were detected by immuno-blotting with m 5 C-specific antibody. Site of cytosine methylation by DR_C0020 encoded protein was investigated by bisulfite sequencing. The DR_C0020 knockout mutant (Δ dcm ), constructed by site directed mutagenesis, was assessed for effect on growth, radiation resistance and proteome. Proteins were identified by mass spectrometry. Results Methylated cytosines were detected in the D. radiodurans genome. The DR_C0020 encoded protein (Dcm, NCBI accession: WP_034351354.1), whose amino acid sequence resembles m 4 C methylases, was shown to be the lone SAM-dependent C-5 cytosine methyltransferase. Purified Dcm protein was found to methylate CpN sequence with a preference for methylation of two consecutive cytosines. The Δ dcm strain completely lost m 5 C modification from its genome, had no effect on growth but became radiation sensitive. The Δ dcm cells exhibited minor alterations in the abundance of several proteins involved primarily in protein homeostasis, oxidative stress defense, metabolism, etc. Conclusion DR_C0020 encoded SAM-dependent methyltransferase Dcm is solely responsible for C-5cytosine methylation at CpN sites in the genome of D. radiodurans and regulates protein homeostasis under normal growth conditions. The protein is an unusual case of an amino methyltransferase that has evolved to producing m 5 C. General significance Although, dispensable under optimal growth conditions, the presence of m 5 C may be important for recognition of parent strand and, thus, could contribute to the extraordinary DNA repair in D. radiodurans . Highlights The genome of D. radiodurans was shown to possess C-5 methylated cytosines. DR_C0020 encoded protein (Dcm) is a SAM-dependent C-5 cytosine methyltransferase. Dcm methylates CpN sites, quite atypical for bacterial methyltransferase. DR_C0020 gene deletion leads to complete loss of m 5 C modification in the genome. Cytosine methylation is dispensable under optimal growth conditions.

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  3. [해외논문]   The promiscuous ectonucleotidase NPP1: molecular insights into substrate binding and hydrolysis   SCI SCIE

    Namasivayam, Vigneshwaran (Corresponding author at: Pharmazeutisches Institut, Pharmazeutische Chemie I, An der Immenburg 4, D-53121 Bonn, Germany.) , Lee, Sang-Yong , Mü , ller, Christa E.
    Biochimica et biophysica acta, General subjects v.1861 no.3 ,pp. 603 - 614 , 2017 , 0304-4165 ,

    초록

    Highlights Substrate preferences of NPP1 were analyzed by combining biochemical experiments, homology modeling and docking studies. All nucleotides are hydrolyzed to nucleoside 5’-monophosphates. The mechanism of 2’,3’’-cGAMP hydrolyis by NPP1 was elucidated. The cyclic dinucleotide 2’,3’’-cGAMP yields 5’-AMP and 5’-GMP, while 3’,3’’-bridged cyclic dinucleotides are not hydrolyzed.

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  4. [해외논문]   Protein toxins of the Echis coloratus viper venom directly activate TRPV1   SCI SCIE

    Geron, Matan (Corresponding author at: Ein Karem Campus, The Hebrew University of Jerusalem, Jerusalem 91120, Israel.) , Kumar, Rakesh , Matzner, Henry , Lahiani, Adi , Gincberg, Galit , Cohen, Gadi , Lazarovici, Philip , Priel, Avi
    Biochimica et biophysica acta, General subjects v.1861 no.3 ,pp. 615 - 623 , 2017 , 0304-4165 ,

    초록

    Abstract Background Peptide and protein toxins are essential tools to dissect and probe the biology of their target receptors. Venoms target vital physiological processes to evoke pain. Snake venoms contain various factors with the ability to evoke, enhance and sustain pain sensation. While a number of venom-derived toxins were shown to directly target TRPV1 channels expressed on somatosensory nerve terminals to evoke pain response, such toxins were yet to be identified in snake venoms. Methods We screened Echis coloratus saw-scaled viper venom's protein fractions isolated by reversed phase HPLC for their ability to activate TRPV1 channels. To this end, we employed heterologous systems to analyze TRPV1 and NGF pathways by imaging and electrophysiology, combined with molecular biology, biochemical, and pharmacological tools. Results We identified TRPV1 activating proteins in the venom of Echis coloratus that produce a channel-dependent increase in intracellular calcium and outwardly rectifying currents in neurons and heterologous systems. Interestingly, channel activation was not mediated by any of its known toxin binding sites. Moreover, although NGF neurotropic activity was detected in this venom, TRPV1 activation was independent of NGF receptors. Conclusions Echis coloratus venom contains proteins with the ability to directly activate TRPV1. This activity is independent of the NGF pathway and is not mediated by known TRPV1 toxins' binding sites. General significance Our results could facilitate the discovery of new toxins targeting TRPV1 to enhance current understanding of this receptor activation mechanism. Furthermore, the findings of this study provide insight into the mechanism through which snakes' venom elicit pain. Highlights The snake venom from Echis coloratus contains peptides that target TRPV1. TRPV1 activation by this venom is not mediated by known toxins' binding sites. Snake venom elicits pain by directly activating pain receptors. The NGF presence in the venom suggests synergistic effect between its components.

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  5. [해외논문]   Revealing the three dimensional architecture of focal adhesion components to explain Ca2+-mediated turnover of focal adhesions   SCI SCIE

    Chang, Shu-Jing (Department of Biomedical Engineering, National Cheng Kung University, Tainan 701, Taiwan ) , Chen, Ying-Chi (Department of Biomedical Engineering, National Cheng Kung University, Tainan 701, Taiwan ) , Yang, Chi-Hsun (Department of Biomedical Engineering, National Cheng Kung University, Tainan 701, Taiwan ) , Huang, Soon-Cen (Department of Obstetrics and Gynecology, Chi Mei Medical Center, Liouying Campus, Tainan 736, Taiwan ) , Huang, Ho-Kai (Department of Biomedical Engineering, National Cheng Kung University, Tainan 701, Taiwan ) , Li, Chun-Chun (Department of Life Sciences, National Cheng Kung University, Tainan 701, Taiwan ) , Harn, Hans I-Chen (Institute of Basic Medical Sciences, National Cheng Kung University, Tainan 701, Taiwan ) , Chiu, Wen-Tai (Department of Biomedical Engineering, National Cheng Kung University, Tainan 701, Taiwan)
    Biochimica et biophysica acta, General subjects v.1861 no.3 ,pp. 624 - 635 , 2017 , 0304-4165 ,

    초록

    Abstract Background Focal adhesions (FAs) are large, dynamic protein complexes located close to the plasma membrane, which serve as the mechanical linkages and a biochemical signaling hub of cells. The coordinated and dynamic regulation of focal adhesion is required for cell migration. Degradation, or turnover, of FAs is a major event at the trailing edge of a migratory cell, and is mediated by Ca 2+ /calpain-dependent proteolysis and disassembly. Here, we investigated how Ca 2+ influx induces cascades of FA turnover in living cells. Methods Images obtained with a total internal reflection fluorescence microscope (TIRFM) showed that Ca 2+ ions induce different processes in the FA molecules focal adhesion kinase (FAK), paxillin, vinculin, and talin. Three mutated calpain-resistant FA molecules, FAK-V744G, paxillin-S95G, and talin-L432G, were used to clarify the role of each FA molecule in FA turnover. Results Vinculin was resistant to degradation and was not significantly affected by the presence of mutated calpain-resistant FA molecules. In contrast, talin was more sensitive to calpain-mediated turnover than the other molecules. Three-dimensional (3D) fluorescence imaging and immunoblotting demonstrated that outer FA molecules were more sensitive to calpain-mediated proteolysis than internal FA molecules. Furthermore, cell contraction is not involved in degradation of FA. Conclusions These results suggest that Ca 2+ -mediated degradation of FAs was mediated by both proteolysis and disassembly. The 3D architecture of FAs is related to the different dynamics of FA molecule degradation during Ca 2+ -mediated FA turnover. General significance This study will help us to clearly understand the underlying mechanism of focal adhesion turnover by Ca 2+ . Highlights Degradation of focal adhesions by Ca 2+ /calpain-dependent proteolysis is demonstrated. Outer FA molecules are more sensitive to calpain-mediated proteolysis. The 3D architecture of FAs is related to the dynamics of FA molecule degradation. Graphical abstract [DISPLAY OMISSION]

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  6. [해외논문]   Structure prediction and network analysis of chitinases from the Cape sundew, Drosera capensis   SCI SCIE

    Unhelkar, Megha H. (Department of Chemistry, University of California, Irvine, Irvine, CA 92697, USA ) , Duong, Vy T. (Department of Chemistry, University of California, Irvine, Irvine, CA 92697, USA ) , Enendu, Kaosoluchi N. (Department of Molecular Biology and Biochemistry, University of California, Irvine, Irvine, CA 92697, USA ) , Kelly, John E. (Department of Chemistry, University of California, Irvine, Irvine, CA 92697, USA ) , Tahir, Seemal (Department of Chemistry, University of California, Irvine, Irvine, CA 92697, USA ) , Butts, Carter T. (Department of Sociology, University of California, Irvine, Irvine, CA 92697, USA ) , Martin, Rachel W. (Department of Chemistry, University of California, Irvine, Irvine, CA 92697, USA)
    Biochimica et biophysica acta, General subjects v.1861 no.3 ,pp. 636 - 643 , 2017 , 0304-4165 ,

    초록

    Abstract Background Carnivorous plants possess diverse sets of enzymes with novel functionalities applicable to biotechnology, proteomics, and bioanalytical research. Chitinases constitute an important class of such enzymes, with future applications including human-safe antifungal agents and pesticides. Here, we compare chitinases from the genome of the carnivorous plant Drosera capensis to those from related carnivorous plants and model organisms. Methods Using comparative modeling, in silico maturation, and molecular dynamics simulation, we produce models of the mature enzymes in aqueous solution. We utilize network analytic techniques to identify similarities and differences in chitinase topology. Results Here, we report molecular models and functional predictions from protein structure networks for eleven new chitinases from D. capensis , including a novel class IV chitinase with two active domains. This architecture has previously been observed in microorganisms but not in plants. We use a combination of comparative and de novo structure prediction followed by molecular dynamics simulation to produce models of the mature forms of these proteins in aqueous solution. Protein structure network analysis of these and other plant chitinases reveal characteristic features of the two major chitinase families. General significance This work demonstrates how computational techniques can facilitate quickly moving from raw sequence data to refined structural models and comparative analysis, and to select promising candidates for subsequent biochemical characterization. This capability is increasingly important given the large and growing body of data from high-throughput genome sequencing, which makes experimental characterization of every target impractical. Highlights We report eleven new chitinases from the carnivorous plant Drosera capensis . A novel two domain class IV chitinase similar to those found in microbes was found. Protein structure prediction and comparison to other carnivorous plant chitinases reveals commonalities. Sequence and structural motifs are conserved among carnivorous plant chitinases. Protein structure networks reveal structural differences and predict functionality. Graphical Abstract [DISPLAY OMISSION]

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  7. [해외논문]   Genomic organization and evolution of olfactory receptors and trace amine-associated receptors in channel catfish, Ictalurus punctatus   SCI SCIE

    Gao, Sen (The Fish Molecular Genetics and Biotechnology Laboratory, Aquatic Genomics Unit, School of Fisheries, Aquaculture and Aquatic Sciences, Auburn University, Auburn, AL 36849, USA ) , Liu, Shikai (The Fish Molecular Genetics and Biotechnology Laboratory, Aquatic Genomics Unit, School of Fisheries, Aquaculture and Aquatic Sciences, Auburn University, Auburn, AL 36849, USA ) , Yao, Jun (The Fish Molecular Genetics and Biotechnology Laboratory, Aquatic Genomics Unit, School of Fisheries, Aquaculture and Aquatic Sciences, Auburn University, Auburn, AL 36849, USA ) , Li, Ning (The Fish Molecular Genetics and Biotechnology Laboratory, Aquatic Genomics Unit, School of Fisheries, Aquaculture and Aquatic Sciences, Auburn University, Auburn, AL 36849, USA ) , Yuan, Zihao (The Fish Molecular Genetics and Biotechnology Laboratory, Aquatic Genomics Unit, School of Fisheries, Aquaculture and Aquatic Sciences, Auburn University, Auburn, AL 36849, USA ) , Zhou, Tao (The Fish Molecular Genetics and Biotechnology Laboratory, Aquatic Genomics Unit, School of Fisheries, Aqu) , Li, Qi , Liu, Zhanjiang
    Biochimica et biophysica acta, General subjects v.1861 no.3 ,pp. 644 - 651 , 2017 , 0304-4165 ,

    초록

    Abstract Background Channel catfish ( Ictalurus punctatus ) live in turbid waters with limited visibility to chase prey within a certain distance. This can be compensated through detecting specific water-soluble substances by the olfactory receptors (ORs) and trace amine associated receptors (TAARs) expressed on the olfactory epithelium. Methods We identified the OR and TAAR repertoires in channel catfish, and characterized the genomic organizations of these two gene families by data mining available genomic resources. Results A total of 47 putative OR genes and 36 putative TAAR genes were identified in the channel catfish genome, including 27 functional OR genes and 28 functional TAAR genes. Phylogenetic and orthogroup analyses were conducted to illustrate the evolutionary dynamics of the vertebrate ORs and TAARs. Collinear analysis revealed the presence of two conserved orthologous blocks that contain OR genes between the catfish genome and zebrafish genome. The complete loss of a conserved motif in fish OR family H may contribute to the divergence of family H from other families. The dN/dS analysis indicated that the highest degree of selection pressure was imposed on TAAR subfamily 14 among all fish ORs and TAARs. Conclusions The present study provides understanding of the evolutionary dynamics of the two gene families (OR and TAAR) associated with olfaction in channel catfish. General significance This is the first systematic study of ORs and TAARs in catfish, which could provide valuable genomic resources for further investigation of olfactory mechanisms in teleost fish. Highlights The OR and TAAR repertoires in channel catfish were identified and characterized by mining existing genomic resources. Two conserved orthologous blocks containing OR genes between the genomes of catfish and zebrafish were identified. The arrangements of conserved motifs within fish ORs and TAARs were generally identical. The complete loss of a conserved motif in fish OR family H may contribute to its divergence from other families. The most selection pressures were imposed on fish TAAR subfamily 14 among all genes involved in olfaction.

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  8. [해외논문]   Proteomic and transcriptomic investigation of acne vulgaris microcystic and papular lesions: Insights in the understanding of its pathophysiology   SCI SCIE

    Quanico, Jusal (Université) , Gimeno, Jean-Pascal (de Lille 1, INSERM U1192 - Laboratoire Protéomique, Réponse Inflammatoire & Spectrométrie de Masse (PRISM), F-59000 Lille, France ) , Nadal-Wollbold, Florence (Université) , Casas, Christiane (de Lille 1, INSERM U1192 - Laboratoire Protéomique, Réponse Inflammatoire & Spectrométrie de Masse (PRISM), F-59000 Lille, France ) , Alvarez-Georges, Sandrine (Centre de Recherche Clinique Pierre-Fabre, Hôtel Dieu, Toulouse, France ) , Redoulè (Centre de Recherche Clinique Pierre-Fabre, Hôtel Dieu, Toulouse, France ) , s, Daniel (Centre de Recherche Clinique Pierre-Fabre, Hôtel Dieu, Toulouse, France ) , Schmitt, Anne-Marie (Centre de Recherche Clinique Pierre-Fabre, Hôtel Dieu, Toulouse, France ) , Fournier, Isabelle (Centre de Recherche Clinique Pierre-Fabre, Hôtel Dieu, Toulouse, France ) , Salzet, Michel (Université)
    Biochimica et biophysica acta, General subjects v.1861 no.3 ,pp. 652 - 663 , 2017 , 0304-4165 ,

    초록

    Abstract Background The pathogenesis of acne vulgaris involves several phases including androgen-dependent hyper-seborrhea, colonization by Propionibacterium acnes , and inflammation. Recent investigations have shown that in fact P. acnes provokes the activation of the inflammasome present in macrophages and dendritic cells. This signaling pathway leads to excessive production of interleukin IL-1β, a proinflammatory cytokine. Nevertheless, these well-studied phenomena in acne fail to elucidate the mechanisms responsible for the appearance of different lesions. Methods We investigate response pathways for specific acne lesions such as microcysts and papules using shot-gun proteomic followed by systemic biology and transcriptomic approaches. Results Results show that most of the proteins identified as differentially expressed between the normal and acne tissue biopsies associated with the immune system response were identified as highly or exclusively expressed in the papule biopsies. They were also expressed in microcysts, but in lower amounts compared to those in papules. These results are supported by the identification of CAMP factor protein produced by P. acnes in microcysts, indicating its enhanced proliferation in this type of lesion Conclusions As CAMP factor protein was not detected in papule biopsies, we can see a clear delineation in the stages of progression of acne pathogenesis, which begins with a hyphenated inflammatory response in the papule stage, followed by imbalance of lipid production, which in turn triggers the enhanced proliferation of P. acnes . General significance We demonstrate that expression inflammation varies across the two types of lesions, suggesting different pathways enhanced as a function of the progression of P. acnes . Highlights Transcriptomic and proteomic approaches of acne vulgaris microcystic and papular lesions Systemic biology studies of metabolic and immune system processes in microcyst and papule Antimicrobial peptides from the acne vulgaris and from microcyst and papule

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  9. [해외논문]   Membrane-assisted viral DNA ejection   SCI SCIE

    Santos-Pé (Structural Biology Unit, CIC bioGUNE, CIBERehd, Bizkaia Technology Park, 48160 Derio, Spain ) , rez, Isaac (Institute of Biotechnology and Department of Biosciences, Viikki Biocenter, University of Helsinki, P.O. Box 56, Viikinkaari 9B, 00014, Finland ) , Oksanen, Hanna M. (Institute of Biotechnology and Department of Biosciences, Viikki Biocenter, University of Helsinki, P.O. Box 56, Viikinkaari 9B, 00014, Finland ) , Bamford, Dennis H. (Unidad de Biofísica (CSIC, UPV/EHU) and Departamento de Bioquímica, Universidad del País Vasco, Bilbao, Spain ) , Goñ (Departament de Física de la Materia Condensada, Facultat de Física, Universitat de Barcelona, 08028 Barcelona, Spain ) , i, Felix M. (Structural Biology Unit, CIC bioGUNE, CIBERehd, Bizkaia Technology Park, 48160 Derio, Spain) , Reguera, David , Abrescia, Nicola G.A.
    Biochimica et biophysica acta, General subjects v.1861 no.3 ,pp. 664 - 672 , 2017 , 0304-4165 ,

    초록

    Abstract Genome packaging and delivery are fundamental steps in the replication cycle of all viruses. Icosahedral viruses with linear double-stranded DNA (dsDNA) usually package their genome into a preformed, rigid procapsid using the power generated by a virus-encoded packaging ATPase. The pressure and stored energy due to this confinement of DNA at a high density is assumed to drive the initial stages of genome ejection. Membrane-containing icosahedral viruses, such as bacteriophage PRD1, present an additional architectural complexity by enclosing their genome within an internal membrane vesicle. Upon adsorption to a host cell, the PRD1 membrane remodels into a proteo-lipidic tube that provides a conduit for passage of the ejected linear dsDNA through the cell envelope. Based on volume analyses of PRD1 membrane vesicles captured by cryo-electron tomography and modeling of the elastic properties of the vesicle, we propose that the internal membrane makes a crucial and active contribution during infection by maintaining the driving force for DNA ejection and countering the internal turgor pressure of the host. These novel functions extend the role of the PRD1 viral membrane beyond tube formation or the mere physical confinement of the genome. The presence and assistance of an internal membrane might constitute a biological advantage that extends also to other viruses that package their linear dsDNA to high density within an internal vesicle. Highlights A semi-empiric approach is layout for studying phage PRD1 internal membrane's role. PRD1 membrane is proposed to actively assist DNA early stages of ejection. PRD1 membrane and assistance might constitute a biological advantage.

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  10. [해외논문]   Hypoxia mimetic induces lipid accumulation through mitochondrial dysfunction and stimulates autophagy in murine preadipocyte cell line   SCI SCIE

    Marques, Ana Patrí (CNC - Center for Neuroscience and Cell Biology, University of Coimbra, 3004-504 Coimbra, Portugal ) , cia (CNC - Center for Neuroscience and Cell Biology, University of Coimbra, 3004-504 Coimbra, Portugal ) , Rosmaninho-Salgado, Joana (CNC - Center for Neuroscience and Cell Biology, University of Coimbra, 3004-504 Coimbra, Portugal ) , Estrada, Marta (CNC - Center for Neuroscience and Cell Biology, University of Coimbra, 3004-504 Coimbra, Portugal ) , Cortez, Vera (CNC - Center for Neuroscience and Cell Biology, University of Coimbra, 3004-504 Coimbra, Portugal ) , Nobre, Rui Jorge (CNC - Center for Neuroscience and Cell Biology, University of Coimbra, 3004-504 Coimbra, Portugal) , Cavadas, Clá , udia
    Biochimica et biophysica acta, General subjects v.1861 no.3 ,pp. 673 - 682 , 2017 , 0304-4165 ,

    초록

    Abstract Background Hypoxia occurs within adipose tissue of obese human and mice. However, its role in adipose tissue regulation is still controversial. Methods We used murine preadipocyte 3T3-L1 cells and hypoxia was induced by using hypoxia mimetic agents, as CoCl 2 . To study adipocyte differentiation, we evaluated the adipocyte markers (PPARγ, C/EBPα and aP2), and a preadipocyte marker (pref-1) by qPCR, western blotting and immunofluorescence. Lipid accumulation was evaluated by Oil red-O assay and perilipin levels by western blotting and immunofluorescence. The effect of CoCl 2 in microRNA, miR-27a and miR-27b, levels was evaluated by qPCR. We also assessed the mitochondrial membrane potential and reactive oxygen species (ROS), superoxide and ATP production. The effect of hypoxia mimetic in autophagy was determined by LC3B and p62 level evaluation by western blotting. Results Our results show that the hypoxia mimetic cobalt chloride increases lipid accumulation with no expression of PPARγ2. Furthermore, using qPCR we observed that the hypoxia mimetic increases microRNAs miR-27a and miR-27b, which are known to block PPARγ2 expression. In contrast, cobalt chloride induces mitochondrial dysfunction, and increases ROS production and autophagy. Moreover, an antioxidant agent, glutathione, prevents lipid accumulation induced by hypoxia mimetic indicating that ROS are responsible for hypoxia-induced lipid accumulation. Conclusions All these results taken together suggest that hypoxia mimetic blocks differentiation and induces autophagy. Hypoxia mimetic also induces lipid accumulation through mitochondrial dysfunction and ROS accumulation. General significance This study highlights the importance of adipocyte response to hypoxia, which might impair adipocyte metabolism and compromise adipose tissue function. Highlights Hypoxia mimetic inhibits adipocyte differentiation and increases miR-27a and b. Hypoxia mimetic causes mitochondrial dysfunction and an increase in reactive oxygen species. Hypoxia mimetic induces lipid accumulation through reactive oxygen species increase. Hypoxia mimetic induces autophagy activation. Graphical abstract [DISPLAY OMISSION]

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