RESUMO
During oogenesis, oocytes gain competence and subsequently undergo meiotic maturation and prepare for embryonic development; trimethylated histone H3 on lysine-4 (H3K4me3) mediates a wide range of nuclear events during these processes. Oocyte-specific knockout of CxxC-finger protein 1 (CXXC1, also known as CFP1) impairs H3K4me3 accumulation and causes changes in chromatin configurations. This study investigated the changes in genomic H3K4me3 landscapes in oocytes with Cxxc1 knockout and the effects on other epigenetic factors such as the DNA methylation, H3K27me3, H2AK119ub1 and H3K36me3. H3K4me3 is overall decreased after knocking out Cxxc1, including both the promoter region and the gene body. CXXC1 and MLL2, which is another histone H3 methyltransferase, have nonoverlapping roles in mediating H3K4 trimethylation during oogenesis. Cxxc1 deletion caused a decrease in DNA methylation levels and affected H3K27me3 and H2AK119ub1 distributions, particularly at regions with high DNA methylation levels. The changes in epigenetic networks implicated by Cxxc1 deletion were correlated with the transcriptional changes in genes in the corresponding genomic regions. This study elucidates the epigenetic changes underlying the phenotypes and molecular defects in oocytes with deleted Cxxc1 and highlights the role of CXXC1 in orchestrating multiple factors that are involved in establishing the appropriate epigenetic states of maternal genome.
Assuntos
Epigênese Genética , Oócitos/metabolismo , Transativadores/fisiologia , Animais , Células Cultivadas , Metilação de DNA , Feminino , Deleção de Genes , Genoma , Código das Histonas , Histonas/metabolismo , Camundongos , Camundongos Endogâmicos C57BL , Regiões Promotoras Genéticas , Transativadores/genética , Transcrição GênicaRESUMO
Stenotrophomonas maltophilia is an endophyte which plays important roles in agricultural production as a plant growth-promoting bacterium. Here, we present the draft genome sequence of strain RR-10, which was isolated from a rice root in a rice field of China.
Assuntos
DNA Bacteriano/química , DNA Bacteriano/genética , Genoma Bacteriano , Oryza/microbiologia , Raízes de Plantas/microbiologia , Stenotrophomonas maltophilia/genética , Stenotrophomonas maltophilia/isolamento & purificação , China , Endófitos/genética , Endófitos/isolamento & purificação , Dados de Sequência Molecular , Análise de Sequência de DNARESUMO
Bombyx mori nucleopolyhedrovirus (BmNPV) ORF54 (Bm54), a member of the viral desmoplakin N-terminus superfamily, is homologous to Autographa californica nucleopolyhedrovirus (AcMNPV) ORF66, which is required for the efficient egress of nucleocapsids from the nucleus and occlusion body formation. In this paper, we generated a bacmid with the Bm54 gene deleted via homologous recombination in Escherichia coli and characterized the mutant virus using a transfection-infection assay and transmission electron microscopy analysis. Our results demonstrated that the cells transfected with viral DNA lacking Bm54 produced non-infectious budded viruses (BVs). Electron microscopy showed that although the deletion of Bm54 did not affect assembly and release of nucleocapsids, it severely affected polyhedron formation. In conclusion, deletion of Bm54 resulted in non-infectious BV and defective polyhedra. Although the sequences of Bm54 and Ac66 are very similar, the two genes function quite differently in the regulation of viral life cycle.
Assuntos
Bombyx/virologia , Desmoplaquinas/metabolismo , Nucleopoliedrovírus/classificação , Nucleopoliedrovírus/genética , Proteínas Virais/metabolismo , Sequência de Aminoácidos , Animais , Anticorpos Antivirais , Bombyx/citologia , Bombyx/ultraestrutura , Células Cultivadas , Desmoplaquinas/genética , Desmoplaquinas/imunologia , Regulação Viral da Expressão Gênica/fisiologia , Microscopia Eletrônica de Transmissão , Nucleopoliedrovírus/fisiologia , Nucleopoliedrovírus/ultraestrutura , Filogenia , Coelhos , Proteínas Virais/genética , Replicação Viral/fisiologiaRESUMO
The Bombyx mori nucleopolyhedrovirus (BmNPV) is a baculovirus that selectively infects domestic silkworm. BmNPV ORF71 (Bm71) is not a core set gene in baculovirus and shares 92 % amino acid sequence identity with Autographa californica multinucleocapsid NPV ORF88 (Ac88/cg30). Previously, it has been reported that virus lacking Ac88 had no striking phenotypes in cell lines or host larvae. However, the exact role of Bm71 during BmNPV life cycle remains unknown. In the present study, we constructed a Bm71-disrupted (Bm71-D) virus and assessed the effect of the Bm71 disruption on viral replication and viral phenotype throughout the viral life cycle. Results showed that the Bm71-D bacmid could successfully transfect Bm5 cell lines and produce infectious budded virus (BV). But the BV titer was 10- to 100-fold lower than that of the wild-type (WT) virus during infection, and the decreased BV titer was rescued by Bm71 gene repair virus (Bm71-R). A larval bioassay showed that Bm71-D virus took 7.5 h longer than the WT to kill Bombyx mori larvae. Transmission electron microscopy analysis indicated that the Bm71-D virus-infected cells had typical virogenic stroma, bundles of nucleocapsids and polyhedra. Taken together, these results suggest that Bm71 has important implications for determining BV yield and virulence in viral life cycle even though it is not an essential gene for replication of BmNPV.
Assuntos
Bombyx/virologia , Nucleopoliedrovírus/patogenicidade , Fases de Leitura Aberta/genética , Liberação de Vírus , Animais , Bioensaio , Bombyx/crescimento & desenvolvimento , Linhagem Celular , Recombinação Homóloga , Larva/virologia , Microscopia Eletrônica de Transmissão , Mutação , Nucleopoliedrovírus/genética , Nucleopoliedrovírus/crescimento & desenvolvimento , Nucleopoliedrovírus/metabolismo , Fases de Leitura Aberta/fisiologia , Transfecção , VirulênciaRESUMO
Pancreatic endocrine islets are vital for glucose homeostasis. However, the islet developmental trajectory and its regulatory network are not well understood. To define the features of these specification and differentiation processes, we isolated individual islet cells from TgBAC(neurod1:EGFP) transgenic zebrafish and analyzed islet developmental dynamics across four different embryonic stages using a single-cell RNA-seq strategy. We identified proliferative endocrine progenitors, which could be further categorized by different cell cycle phases with the G1/S subpopulation displaying a distinct differentiation potential. We identified endocrine precursors, a heterogeneous intermediate-state population consisting of lineage-primed alpha, beta and delta cells that were characterized by the expression of lineage-specific transcription factors and relatively low expression of terminally differentiation markers. The terminally differentiated alpha, beta, and delta cells displayed stage-dependent differentiation states, which were related to their functional maturation. Our data unveiled distinct states, events and molecular features during the islet developmental transition, and provided resources to comprehensively understand the lineage hierarchy of islet development at the single-cell level.
Assuntos
Antígenos de Diferenciação/biossíntese , Regulação da Expressão Gênica no Desenvolvimento , Ilhotas Pancreáticas/embriologia , Análise de Célula Única , Transcrição Gênica , Peixe-Zebra/embriologia , Animais , Animais Geneticamente Modificados/embriologia , Animais Geneticamente Modificados/genética , Antígenos de Diferenciação/genética , Peixe-Zebra/genéticaRESUMO
Two novel Wells-Dawson polyoxometalate-based compounds, (H2bimb)2[{Ag1.5(bimb)1.5}(P2W18O62)] (1), (H2bimb)2[Ag(bimb)]0.5[Ag(eim)2]0.5[P2W18O62]·H3O (2) (bimb = 1,4-bis(imidazole-l-yl)butane, eim = ethyl(imidazole)), have been synthesized under hydrothermal conditions and characterized by IR, thermogravimetric, XRD and single-crystal X-ray diffraction analyses. In compound 1, two "U"-type bimb ligands are connected together viaπ interactions of Ag-C to form a ring unit. The ring unit alternately links with the "Z"-type bimb via an Ag-N bond to form an infinite Z-shaped {Ag4(bimb)3}n chain. The adjacent two {P2W18} clusters bonded to two Ag1 ions on two adjacent Z-shaped chains to form an inorganic dimer linker, which further joins the Z-shaped chains together, resulting in unique organic-inorganic alternating 2-D layers. Adjacent 2D layers are further aggregated together via Ag-O bonds between the Dawson cluster and the Z-type chain on different layers to yield a 3-D network with a new topology {103}2{10}{4·102}4{42·104}2. In compound 2, the eim ligand was synthesized in situ from bimb, which acts as a small molecular ligand to alter the structure and adjust the catalytic activity of compound 2. Each Ag2 connects with the bimb ligand to form a linear {Ag(bimb)} chain. Adjacent chains are bonded together via the bimb ligand to generate a supermolecule 2-D layer, in which a Dawson dimer cluster as a guest molecule is embedded in both sides of the layer through supermolecule interactions. Such 2-D layers are further supported by Ag(eim)2 units leading to 3D supermolecule nets. In addition, the compounds exhibit excellent electrocatalytic behavior and better photocatalytic degradation capability for typical dyes than other Wells-Dawson POMs.