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A diatom-associated bacterium, designated as strain F10T, was isolated from a pure culture of the pennate diatom Asterionellopsis glacialis A3 and has since been used to characterize molecular mechanisms of symbiosis between phytoplankton and bacteria, including interactions using diatom-derived azelaic acid. Its origin from a hypersaline environment, combined with its capacity for quorum sensing, biofilm formation, and potential for dimethylsulfoniopropionate methylation/cleavage, suggest it is within the family Roseobacteraceae. Initial phylogenetic analysis of the 16S rRNA gene sequence placed this isolate within the Phaeobacter genus, but recent genomic and phylogenomic analyses show strain F10T is a separate lineage diverging from the genus Pseudophaeobacter. The genomic DNA G+C content is 60.0 mol%. The predominant respiratory quinone is Q-10. The major fatty acids are C18â:â1 ω7c and C16â:â0. Strain F10T also contains C10â:â03-OH and the furan-containing fatty acid 10,13-epoxy-11-methyl-octadecadienoate (9-(3-methyl-5-pentylfuran-2-yl)nonanoic acid). The major polar lipids are diphosphatidylglycerol, phosphatidylethanolamine and phosphatidylglycerol. Based on genomic, phylogenomic, phenotypic and chemotaxonomic characterizations, strain F10T represents a novel genus and species with the proposed name, Phycobacter azelaicus gen. nov. sp. nov. The type strain is F10T (=NCMA B37T=NCIMB 15470T=NRIC 2002T).
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Diatomáceas , Rhodobacteraceae , Ácidos Graxos/química , Fosfolipídeos/análise , Diatomáceas/genética , Ubiquinona , Filogenia , RNA Ribossômico 16S/genética , Análise de Sequência de DNA , DNA Bacteriano/genética , Composição de Bases , Técnicas de Tipagem Bacteriana , Rhodobacteraceae/genéticaRESUMO
Unicellular eukaryotic phytoplankton, such as diatoms, rely on microbial communities for survival despite lacking specialized compartments to house microbiomes (e.g., animal gut). Microbial communities have been widely shown to benefit from diatom excretions that accumulate within the microenvironment surrounding phytoplankton cells, known as the phycosphere. However, mechanisms that enable diatoms and other unicellular eukaryotes to nurture specific microbiomes by fostering beneficial bacteria and repelling harmful ones are mostly unknown. We hypothesized that diatom exudates may tune microbial communities and employed an integrated multiomics approach using the ubiquitous diatom Asterionellopsis glacialis to reveal how it modulates its naturally associated bacteria. We show that A. glacialis reprograms its transcriptional and metabolic profiles in response to bacteria to secrete a suite of central metabolites and two unusual secondary metabolites, rosmarinic acid and azelaic acid. While central metabolites are utilized by potential bacterial symbionts and opportunists alike, rosmarinic acid promotes attachment of beneficial bacteria to the diatom and simultaneously suppresses the attachment of opportunists. Similarly, azelaic acid enhances growth of beneficial bacteria while simultaneously inhibiting growth of opportunistic ones. We further show that the bacterial response to azelaic acid is numerically rare but globally distributed in the world's oceans and taxonomically restricted to a handful of bacterial genera. Our results demonstrate the innate ability of an important unicellular eukaryotic group to modulate select bacteria in their microbial consortia, similar to higher eukaryotes, using unique secondary metabolites that regulate bacterial growth and behavior inversely across different bacterial populations.
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Bactérias/crescimento & desenvolvimento , Diatomáceas/metabolismo , Microbiota/fisiologia , Fitoplâncton/metabolismo , Microbiologia da Água , Animais , Bactérias/genética , Cinamatos/metabolismo , Depsídeos/metabolismo , Diatomáceas/genética , Ácidos Dicarboxílicos/metabolismo , Perfilação da Expressão Gênica , Metabolômica , Metagenoma , Metagenômica , Oceanos e Mares , Fitoplâncton/genética , Metabolismo Secundário/fisiologia , Ácido RosmarínicoRESUMO
Interactions between phytoplankton and bacteria play major roles in global biogeochemical cycles and oceanic nutrient fluxes. These interactions occur in the microenvironment surrounding phytoplankton cells, known as the phycosphere. Bacteria in the phycosphere use either chemotaxis or attachment to benefit from algal excretions. Both processes are regulated by quorum sensing (QS), a cell-cell signalling mechanism that uses small infochemicals to coordinate bacterial gene expression. However, the role of QS in regulating bacterial attachment in the phycosphere is not clear. Here, we isolated a Sulfitobacter pseudonitzschiae F5 and a Phaeobacter sp. F10 belonging to the marine Roseobacter group and an Alteromonas macleodii F12 belonging to Alteromonadaceae, from the microbial community of the ubiquitous diatom Asterionellopsis glacialis. We show that only the Roseobacter group isolates (diatom symbionts) can attach to diatom transparent exopolymeric particles. Despite all three bacteria possessing genes involved in motility, chemotaxis, and attachment, only S. pseudonitzschiae F5 and Phaeobacter sp. F10 possessed complete QS systems and could synthesize QS signals. Using UHPLC-MS/MS, we identified three QS molecules produced by both bacteria of which only 3-oxo-C16:1 -HSL strongly inhibited bacterial motility and stimulated attachment in the phycosphere. These findings suggest that QS signals enable colonization of the phycosphere by algal symbionts.
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Aderência Bacteriana , Diatomáceas/microbiologia , Locomoção , Fitoplâncton/microbiologia , Percepção de Quorum/fisiologia , 4-Butirolactona/análogos & derivados , 4-Butirolactona/metabolismo , Bactérias/classificação , Bactérias/genética , Bactérias/metabolismo , Aderência Bacteriana/genética , Genes Bacterianos , Locomoção/genética , Microbiota , Oceanos e Mares , Percepção de Quorum/genéticaRESUMO
Low-density lipoprotein receptor-related proteins 5 and 6 (LRP5/6) are co-receptors for Wnt ligands. Upon ligand binding, LRP5/6 undergo glycogen synthase kinase 3 (GSK3)/casein kinase I (CKI)-mediated phosphorylation at multiple PPP(S/T)P motifs in the intracellular domain, which is essential for canonical Wnt signal transduction. On the other hand, in the Wnt-off state, the mitosis-specific CDK14-Cyclin Y kinase complex phosphorylates Ser-1490 of LRP5/6 at G2/M, thereby priming the receptor for Wnt-induced phosphorylation. However, it remains unclear how CDK14/Cyclin Y is recruited to LRP5/6 and whether there are other cofactors involved in this process. Previously, we identified Caprin-2 as a positive regulator of canonical Wnt signaling by promoting GSK3-depedent LRP5/6 phosphorylation upon Wnt stimulation. Here we uncovered that Caprin-2 positively regulates constitutive LRP5/6 Ser-1490 phosphorylation by complexing with CDK14/Cyclin Y. Caprin-2-mediated LRP5/6 phosphorylation is cell cycle-dependent in a pattern similar to that of CDK14/Cyclin Y-dependent LRP5/6 phosphorylation. Moreover, knockdown of Caprin-2 disrupts not only the interaction between CDK14 and Cyclin Y but also the interaction between CDK14/Cyclin Y and LRP6. Overall, our findings revealed an unrecognized role of Caprin-2 in facilitating LRP5/6 constitutive phosphorylation at G2/M through forming a quaternary complex with CDK14, Cyclin Y, and LRP5/6.
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Proteínas de Ciclo Celular/metabolismo , Divisão Celular/fisiologia , Quinases Ciclina-Dependentes/metabolismo , Ciclinas/metabolismo , Fase G2/fisiologia , Proteína-5 Relacionada a Receptor de Lipoproteína de Baixa Densidade/metabolismo , Proteína-6 Relacionada a Receptor de Lipoproteína de Baixa Densidade/metabolismo , Proteínas de Ciclo Celular/genética , Quinases Ciclina-Dependentes/genética , Ciclinas/genética , Técnicas de Silenciamento de Genes , Células HEK293 , Células HeLa , Humanos , Proteína-5 Relacionada a Receptor de Lipoproteína de Baixa Densidade/genética , Proteína-6 Relacionada a Receptor de Lipoproteína de Baixa Densidade/genética , Fosforilação/fisiologia , Proteínas de Ligação a RNA , Via de Sinalização Wnt/fisiologiaRESUMO
Previously, Smad ubiquitination regulatory factor 1 (Smurf1)-mediated Lys29 (K29)-linked poly-ubiquitination of Axin has been identified as a novel regulatory process in Wnt/ß-catenin signaling. In this work, we discovered that the C2 domain of Smurf1 is critical for targeting Axin for ubiquitination. We found that the C2 domain-mediated plasma membrane localization of Smurf1 is required for Axin ubiquitination, and interfering with that disturbs the co-localization of Smurf1 and Axin around the plasma membrane. Moreover, the C2 domain of Smurf1, rather than its WW domains, is involved in Smurf1's interaction with Axin; and the putative PPXY motifs (PY motif) of Axin are not essential for such an interaction, indicating that Smurf1 binds to Axin in a non-canonical way independent of WW-PY interaction. Further, we found that Smurf1-Axin interaction and Axin ubiquitination are attenuated in the G2/M phase of cell cycle, contributing to an increased cell response to Wnt stimulation at that stage. Collectively, we uncovered a dual role of Smurf1 C2 domain, recruiting Smurf1 to membrane for accessing Axin and mediating its interaction with Axin, and that Smurf1-mediated Axin ubiquitination is subjected to the regulation of cell cycle.
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Proteína Axina/metabolismo , Ciclo Celular , Ubiquitina-Proteína Ligases/química , Ubiquitina-Proteína Ligases/metabolismo , Ubiquitinação , Animais , Linhagem Celular , Membrana Celular/metabolismo , Humanos , Ligação Proteica , Estrutura Terciária de Proteína , Transporte Proteico , Via de Sinalização Wnt , beta Catenina/metabolismoRESUMO
The Wnt/ß-catenin signaling pathway has a crucial role in embryonic development, stem cell maintenance and human disease. By screening a synthetic chemical library of lycorine derivatives, we identified 4-ethyl-5-methyl-5,6-dihydro-[1,3]dioxolo[4,5-j]phenanthridine (HLY78) as an activator of the Wnt/ß-catenin signaling pathway, which acts in a Wnt ligand-dependent manner. HLY78 targets the DIX domain of Axin and potentiates the Axin-LRP6 association, thus promoting LRP6 phosphorylation and Wnt signaling transduction. Moreover, we identified the critical residues on Axin for HLY78 binding and showed that HLY78 may weaken the autoinhibition of Axin. In addition, HLY78 acts synergistically with Wnt in the embryonic development of zebrafish and increases the expression of the conserved hematopoietic stem cell (HSC) markers, runx1 and cmyb, in zebrafish embryos. Collectively, our study not only provides new insights into the regulation of the Wnt/ß-catenin signaling pathway by a Wnt-specific small molecule but also will facilitate therapeutic applications, such as HSC expansion.
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Proteína Axina/metabolismo , Benzodioxóis/farmacologia , Proteína-5 Relacionada a Receptor de Lipoproteína de Baixa Densidade/metabolismo , Proteína-6 Relacionada a Receptor de Lipoproteína de Baixa Densidade/metabolismo , Fenantridinas/farmacologia , Bibliotecas de Moléculas Pequenas/farmacologia , Proteínas Wnt/metabolismo , Via de Sinalização Wnt/efeitos dos fármacos , Animais , Proteína Axina/antagonistas & inibidores , Proteína Axina/química , Benzodioxóis/química , Células HEK293 , Humanos , Proteína-5 Relacionada a Receptor de Lipoproteína de Baixa Densidade/química , Proteína-6 Relacionada a Receptor de Lipoproteína de Baixa Densidade/química , Fenantridinas/química , Ligação Proteica/efeitos dos fármacos , Bibliotecas de Moléculas Pequenas/química , Peixe-Zebra/embriologia , Peixe-Zebra/metabolismoRESUMO
Cotton is a significant cash crop and the primary source of natural fiber globally. Among the numerous diseases encountered in cotton production, Verticillium wilt is one of the most serious, caused by the pathogen Verticillium dahliae (V. dahliae). Unfortunately, there are no effective targeted methods to combat this disease. Genomic resources for Verticillium wilt resistance primarily exist in Gossypium barbadense (G. barbadense). Regrettably, there have been limited transcriptomic comparisons between V. dahliae-resistant and -susceptible varieties of G. barbadense due to the scarcity of susceptible resources. In this study, we conducted a transcriptome analysis on both V. dahliae-resistant and -susceptible varieties of G. barbadense at the 0, 12, 24 and 48 hours after V. dahliae inoculation. This comparative transcriptome analysis yielded high-quality data and offered new insights into the molecular mechanisms underlying cotton's resistance against this destructive pathogen.
Assuntos
Gossypium , Verticillium , Resistência à Doença/genética , Perfilação da Expressão Gênica , Regulação da Expressão Gênica de Plantas , Gossypium/genética , Doenças das Plantas/genética , Verticillium/genéticaRESUMO
Interactions between phytoplankton and bacteria play important roles in shaping the microenvironment surrounding these organisms and in turn influence global biogeochemical cycles. This microenvironment, known as the phycosphere, is presumed to shape the bacterial diversity around phytoplankton and thus stimulate a diverse array of interactions between both groups. Although many studies have attempted to characterize bacterial communities that associate and interact with phytoplankton, bias in bacterial cultivation and consistency and persistence of bacterial communities across phytoplankton isolates likely impede the understanding of these microbial associations. Here, we isolate four strains of the diatom Asterionellopsis glacialis and three strains of the diatom Nitzschia longissima and show through metabarcoding of the bacterial 16S rDNA gene that though each species possesses a unique bacterial community, the bacterial composition across strains from the same species are highly conserved at the genus level. Cultivation of all seven strains in the laboratory for longer than 1 year resulted in only small changes to the bacterial composition, suggesting that despite strong pressures from laboratory culturing conditions associations between these diatoms and their bacterial communities are robust. Specific operational taxonomic units (OTUs) belonging to the Roseobacter-clade appear to be conserved across all strains and time, suggesting their importance to diatoms. In addition, we isolate a range of cultivable bacteria from one of these cultures, A. glacialis strain A3, including several strains of Shimia marina and Nautella sp. that appear closely related to OTUs conserved across all strains and times. Coculturing of A3 with some of its cultivable bacteria as well as other diatom-associated bacteria shows a wide range of responses that include enhancing diatom growth. Cumulatively, these findings suggest that phytoplankton possess unique microbiomes that are consistent across strains and temporal scales.
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Wnt/ß-catenin signaling regulates multiple biological processes and aberration of this pathway is frequently observed in human cancers. Previously, we uncovered NC043 as a small-molecule inhibitor of Wnt/ß-catenin signaling. Here, we identified CARF as the cellular target of NC043. We found that NC043 binds directly to CARF through forming a covalent bond with the Cys-516 residue of CARF. Further study revealed that CARF interacts with Dvl, which potentiates the Dvl-c-Jun-ß-catenin-TCF transcriptional complex and thus promotes Wnt signaling activation. NC043 could disrupt the interaction between CARF and Dvl, thereby impairing Wnt signal transduction. In line with this, knockdown of CARF in zebrafish leads to impairment of embryonic development, hematopoietic stem cell generation and caudal fin regeneration. Collectively, we identified CARF as the cellular target of NC043 and revealed CARF as a positive regulator of Wnt/ß-catenin signal transduction.
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Limbs’soft tissue injury is complex,often accompanied by vascular injury. Improper treatment will bring disastrous consequences including serious disability even amputation. Numerous evaluation criteria are available for limbs’soft tissue injury,but there is no unified standard. Accurate injury assessment is not only the basis of treatment but also a clinical difficulty. Treatment regimes are controversial,for example,different opinions on repair at stage I or stage II and on repair using traditional stair-step method or perforator flap when treating the same injury. The authors have a discussion on soft tissue injury with regards to its characteristics and assessment,treatment concept and treatment points,so as to provide references for the treatment of limbs’ soft tissue injury.
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Several different barcoding methods of distinguishing species have been advanced, but which method is the best is still controversial. Chlorella is becoming particularly promising in the development of second-generation biofuels. However, the taxonomy of Chlorella-like organisms is easily confused. Here we report a comprehensive barcoding analysis of Chlorella-like species from Chlorella, Chloroidium, Dictyosphaerium and Actinastrum based on rbcL, ITS, tufA and 16S sequences to test the efficiency of traditional barcoding, GMYC, ABGD, PTP, P ID and character-based barcoding methods. First of all, the barcoding results gave new insights into the taxonomic assessment of Chlorella-like organisms studied, including the clear species discrimination and resolution of potentially cryptic species complexes in C. sorokiniana, D. ehrenbergianum and C. Vulgaris. The tufA proved to be the most efficient barcoding locus, which thus could be as potential "specific barcode" for Chlorella-like species. The 16S failed in discriminating most closely related species. The resolution of GMYC, PTP, P ID, ABGD and character-based barcoding methods were variable among rbcL, ITS and tufA genes. The best resolution for species differentiation appeared in tufA analysis where GMYC, PTP, ABGD and character-based approaches produced consistent groups while the PTP method over-split the taxa. The character analysis of rbcL, ITS and tufA sequences could clearly distinguish all taxonomic groups respectively, including the potentially cryptic lineages, with many character attributes. Thus, the character-based barcoding provides an attractive complement to coalescent and distance-based barcoding. Our study represents the test that proves the efficiency of multiple DNA barcoding in species discrimination of microalgaes.
Assuntos
Chlorella/classificação , Chlorella/genética , Microalgas/classificação , Microalgas/genética , Código de Barras de DNA Taxonômico/métodos , FilogeniaRESUMO
Sugar beet (Beta vulgaris cv. Beta 356) was subjected to drought stress during vegetative development by maintaining the soil water content in the 0-40 cm soil depth at 70%, 50% or 30% of field capacity to study the physiological traits of the leaves. Results showed that the compensation index was the highest in the 50% field capacity treatment. Malonaldehyde (MDA) content, relative conductivity, catalase (CAT) activity, and soluble sugar content began to increase 24 h after rehydration. Proline content began to increase 48 h after rehydration. In contrast, no compensation effect was observed in peroxidase (POD) activity after rehydration. Among the active oxygen scavenging enzymes, CAT was most sensitive to drought stress. Supplemental irrigation should be carried out promptly when the soil water content dropped to 50% of field capacity during vegetative development. Rehydration could promote self-repair functions in leaves, thus reducing the effects of drought on sugar beet yield and sugar content.
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Irrigação Agrícola , Beta vulgaris/fisiologia , Secas , Estresse Fisiológico , Água/fisiologia , Carboidratos/química , Catalase/química , Malondialdeído/química , Peroxidases/química , Folhas de Planta/fisiologia , Prolina/química , SoloRESUMO
Microalgae identification is extremely difficult. The efficiency of DNA barcoding in microalgae identification involves ideal gene markers and approaches employed, which however, is still under the way. Although Scenedesmus has obtained much research in producing lipids its identification is difficult. Here we present a comprehensive coalescent, distance and character-based DNA barcoding for 118 Scenedesmus strains based on rbcL, tufA, ITS and 16S. The four genes, and their combined data rbcL + tufA + ITS + 16S, rbcL + tufA and ITS + 16S were analyzed by all of GMYC, P ID, PTP, ABGD, and character-based barcoding respectively. It was apparent that the three combined gene data showed a higher proportion of resolution success than the single gene. In comparison, the GMYC and PTP analysis produced more taxonomic lineages. The ABGD generated various resolution in discrimination among the single and combined data. The character-based barcoding was proved to be the most effective approach for species discrimination in both single and combined data which produced consistent species identification. All the integrated results recovered 11 species, five out of which were revealed as potential cryptic species. We suggest that the character-based DNA barcoding together with other approaches based on multiple genes and their combined data could be more effective in microalgae diversity revelation.
Assuntos
Código de Barras de DNA Taxonômico/métodos , Proteínas de Plantas/genética , Scenedesmus/classificação , DNA Ribossômico/genética , Filogenia , Scenedesmus/genética , Especificidade da EspécieRESUMO
It was economically feasible to screen strains adaptive to wide temperature fluctuation for outdoor cultivation without temperature control. In this research, three Chlorella strains from arctic glacier, desert soil and temperate native lake were isolated and identified. The growth, biochemical composition, lipid content and fatty acid composition of each strain cultured under the mode of diurnal temperature fluctuations were compared. All the three Chlorella strains showed desirable abilities of accumulating lipid under diurnal temperature fluctuations and their fatty acid profiles were suitable for biodiesel production, although the growth and biochemical composition were seemed to be region-specific. The highest lipid content was at 51.83±2.49% DW, 42.80±2.97% DW and 36.13±2.27% DW under different temperature fluctuation of 11 °C, 25 °C, 7 °C, respectively. The results indicated that the three Chlorella strains could be promising biodiesel feedstock for outdoor cultivation by the cultural mode of diurnal temperature fluctuations.
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Chlorella/crescimento & desenvolvimento , Chlorella/metabolismo , Ritmo Circadiano , Lipídeos/biossíntese , Temperatura , Proteínas de Algas/análise , Regiões Árticas , Biocombustíveis , Biomassa , Chlorella/isolamento & purificação , Clorofila/análise , Ácidos Graxos/biossíntese , FilogeniaRESUMO
To improve the biomass yield and lipid productivity, two desert microalgae, Desmodesmus sp. S81 and G41 were induced mutagenesis by Ethylmethane sulfonate (EMS), and obtained two potential mutants, Desmodesmus sp. S5 and G3 from the mutagenic clones for their greatly promoted biomass and lipid production. The results showed that the biomass yield, lipid content and lipid productivity of the mutant strains S5 and G3 were 778.10mg·L(-1), 48.41% and 19.83mg·L(-1)·d(-1), 739.52mg·L(-1), 46.01%, and 17.92mg·L(-1)·d(-1), respectively, which presented the increment of 45.50%, 8.00% and 74.24%, 20.67%, 10.35% and 55.77% than those of S81 and G41. Comparing with the wild strains, the mutants showed reduced PUFAs and glycol lipids, elevated MUFAs and neutral lipids contents, which were appropriate for biodiesel production.
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Biomassa , Técnicas de Cultura de Células/métodos , Clorófitas/crescimento & desenvolvimento , Clorófitas/genética , Metanossulfonato de Etila/farmacologia , Lipídeos/biossíntese , Mutação/genética , Biocombustíveis/microbiologia , Clorofila/metabolismo , Clorófitas/efeitos dos fármacos , Clorófitas/metabolismo , Ácidos Graxos/metabolismo , Fluorescência , Microalgas/genética , Microalgas/crescimento & desenvolvimento , Microalgas/metabolismo , Mutagênese , Oxazinas/metabolismo , Fatores de TempoRESUMO
Aim To explore whether histone modifications are involved in the process of uterine injury alleviated by TSA in female mice induced by cigarette smoke (CS) exposure. Methods Female mice were exposed to CS twice daily for 30 days and TSA was injected intraperitoneally into CS-exposed mice on alternate days in the TSA-treated group. HE staining was used to observe the morphological changes of mice uterus after CS exposure; Western blot was used to assess the global modification levels of H3K4mel, H3K4me2, H3K4me3, H3K9mel, H3K9me2, H3K9me3 and H3K27me3 in uteri. GLP (H3K9 his-tone methyltransferase) , G9a (H3K9 histone methyl- transferase) , EZH2 (H3K27me3 histone methyltrans ferase ). Results TSA effectively restored the number of glandular and interstitial cells reduced by CS exposure. Western blot results showed that TSA significantly inhibited global H3K9mel modification and further aggravated H3K27me3 change induced by CS exposure. Furthermore TSA suppressed GLP and G9n expression in mouse uterine tissue induced by CS exposure, but further activated EZH2 increase. Conclusions Histone modifications are involved in the process of uterine injury alleviated by TSA in female mice induced by cigarette smoke exposure.
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Ubiquitination plays important and diverse roles in modulating protein functions. As a C2-WW-HECT-type ubiquitin ligase, Smad ubiquitination regulatory factor 1 (Smurf1) commonly serves to regulate ubiquitin-dependent protein degradation in a number of signaling pathways. Here, we report a novel function of Smurf1 in regulating Wnt/ß-catenin signaling through targeting axin for nonproteolytic ubiquitination. Our data unambiguously demonstrate that Smurf1 ubiquitinates axin through Lys 29 (K29)-linked polyubiquitin chains. Unexpectedly, Smurf1-mediated axin ubiquitination does not lead to its degradation but instead disrupts its interaction with the Wnt coreceptors LRP5/6, which subsequently attenuates Wnt-stimulated LRP6 phosphorylation and represses Wnt/ß-catenin signaling. The inhibitory function of Smurf1 on Wnt/ß-catenin signaling is further evidenced by analysis with Smurf1 knockout murine embryonic fibroblasts. We next identified K789 and K821 in axin as the ubiquitination sites by Smurf1. Consistently, Smurf1 could neither disrupt the interaction of an axin(K789/821R) double mutant with LRP5/6 nor attenuate the phosphorylation of LRP6 in axin(K789/821R)-expressing cells. Collectively, our studies uncover Smurf1 as a new regulator for the Wnt/ß-catenin signaling pathway via modulating the activity of axin.
Assuntos
Proteína Axina/metabolismo , Lisina/metabolismo , Transdução de Sinais , Ubiquitina-Proteína Ligases/metabolismo , Ubiquitina/metabolismo , Proteínas Wnt/metabolismo , beta Catenina/metabolismo , Animais , Proteína Axina/genética , Fibroblastos/citologia , Fibroblastos/metabolismo , Regulação da Expressão Gênica , Células HEK293 , Humanos , Proteína-5 Relacionada a Receptor de Lipoproteína de Baixa Densidade/genética , Proteína-5 Relacionada a Receptor de Lipoproteína de Baixa Densidade/metabolismo , Proteína-6 Relacionada a Receptor de Lipoproteína de Baixa Densidade/genética , Proteína-6 Relacionada a Receptor de Lipoproteína de Baixa Densidade/metabolismo , Lisina/genética , Camundongos , Mutação , Fosforilação , Ubiquitina/genética , Ubiquitina-Proteína Ligases/genética , Ubiquitinação , Proteínas Wnt/genética , beta Catenina/genéticaRESUMO
Objective To explore the method of limb salvage therapy for hand and foot destructive injury, and to clarify the role of free tissue transplantation in limb salvage therapy in patients with hand and foot destructive injury. Methods From January, 2015 to January, 2016, 17 hand and foot destructive injuried patients were treated with limb salvage. The anterolateral thigh free flap, free fibular flap and arterial vein flap were used to repair the skin, soft tissue and bone defects. All patients were followed-up for 12 months. The appearance and function of the patients were evalu-ated after the treatment. The hand scoring criterion was the finger replantation function evaluation standard on probation of the Chinese Medical Association of Hand Surgery. The foot scoring criterion was the Maryland score system. Re-sults All flaps survived in 17 patients. One flap had vascular crisis after operation, and survived after vascular ex-ploratory operation. One case had infection postoperation and healed after several debridements. After 12 months, the functional scores were excellent in 5 cases, good in 5 cases, moderate in 3 cases and poor in 4 cases. Conclusion The early repair with free tissue transplantation can preserve limb length, cover the exposed bone and tendon, and improve the anti-infective ability, so as to improve the function of limb salvage and patient 's satisfaction.
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Objective To explore the curative effect of fibular flap with limbs composite soft-tissue.Methods From February,2013 to February,2016,13 cases with body severe trauma patients were treated,which including 5 cases of upper limbs and 8 cases of lower limbs,and all existed bone defect,soft tissue defect and trunk vessel defect.Three cases with limbs distal non blood supply were emergency treated with debridment and flow-through fibular flap transplantation renovation,peroneal artery repairing defective blood vesscls to rcstorc limbs distal blood supply,fibular flap repairing bone defect,skin flap repairing soft tissue defect.The limb blood supply for other 10 cases were in good condition,but one case with main artery defect did the second phase of fibular flap transplantation and repaired defective blood vessels,bone and skin soft tissue synchronously according to wound condition.According to the postoperative observation for flap survival and appearance,X-ray films to observe fracture healing after 6 weeks,three months and 6 months of operation as well as evaluating limb function recovery,then analyzed the results.Results Flaps survived successfully for 11 cases,and flaps for the other 2 cases were partial necrosis.One Case was edge flap necrosis,heal scabby after dressing,and the other case was necrosis for 1/3 of the area,but the deep fascia survival,and the skin graft healing after dressing.One case with forearm rolling was in vascular crisis after operation,but tbe crisis was relieved after detection,and fingers blood supply was recovered.All the patients were followed up for 6 to 36 months(mean,14 months).All flaps were survived,fractures healed well and limbs distal blood supply was good.Bone healing time was 8 to 24 weeks,and patients with lower limbs injury could bear load after 3 to 8 months.Lower limbs restored walking function.Upper limbs and hands restored rotation function.Transplant flapshad good elasticity and satisfactory appearance.Conclusion Using fibular flap to repair defective blood vessels,bone and soft tissue synchronously,not only can rescue the limbs on the verge of amputation,but also can repair defective composite tissue and get a good prognosis.It is an effective method for open injuries severely treatment in clinic.
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Members of the bone morphogenetic protein (BMP) and T-box gene families play several critical roles in the early embryonic development and tissue homeostasis. Although BMP proteins are the upstream regulators of T-box genes, few studies have investigated the molecular mechanisms between these two protein families. Here, we report that Tbx6 interacts directly with Smad6, an inhibitory Smad that antagonizes the BMP signal. This interaction is mediated through the Mad homology 2 (MH2) domain of Smad6 and residues 90-180 of Tbx6. We demonstrate that Smad6 facilitates the degradation of Tbx6 protein through recruitment of Smurf1, a ubiquitin E3 ligase. Consequently, Smad6 reduces Tbx6-mediated Myf-5 gene activation. Furthermore, specific knockdown of endogenous Smad6 and Smurf1 by small interfering RNA increases the protein levels of Tbx6 and enhance the expression of Tbx6 target genes. Collectively, these findings reveal that Smad6 serves as a critical mediator of BMP signal via a functional interaction with Tbx6, thus regulating the activation of Tbx6 downstream genes during cell differentiation.