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Epstein-Barr virus (EBV) BKRF3 shares sequence homology with members of the uracil-N-glycosylase (UNG) protein family and has DNA glycosylase activity. Here, we explored how BKRF3 participates in the DNA replication complex and contributes to viral DNA replication. Exogenously expressed Flag-BKRF3 was distributed mostly in the cytoplasm, whereas BKRF3 was translocated into the nucleus and colocalized with the EBV DNA polymerase BALF5 in the replication compartment during EBV lytic replication. The expression level of BKRF3 increased gradually during viral replication, coupled with a decrease of cellular UNG2, suggesting BKRF3 enzyme activity compensates for UNG2 and ensures the fidelity of viral DNA replication. In immunoprecipitation-Western blotting, BKRF3 was coimmunoprecipitated with BALF5, the polymerase processivity factor BMRF1, and the immediate-early transactivator Rta. Coexpression of BMRF1 appeared to facilitate the nuclear targeting of BKRF3 in immunofluorescence staining. Residues 164 to 255 of BKRF3 were required for interaction with Rta and BALF5, whereas residues 81 to 166 of BKRF3 were critical for BMRF1 interaction in glutathione S-transferase (GST) pulldown experiments. Viral DNA replication was defective in cells harboring BKRF3 knockout EBV bacmids. In complementation assays, the catalytic mutant BKRF3(Q90L,D91N) restored viral DNA replication, whereas the leucine loop mutant BKRF3(H213L) only partially rescued viral DNA replication, coupled with a reduced ability to interact with the viral DNA polymerase and Rta. Our data suggest that BKRF3 plays a critical role in viral DNA synthesis predominantly through its interactions with viral proteins in the DNA replication compartment, while its enzymatic activity may be supplementary for uracil DNA glycosylase (UDG) function during virus replication. IMPORTANCECatalytic activities of both cellular UDG UNG2 and viral UDGs contribute to herpesviral DNA replication. To ensure that the enzyme activity executes at the right time and the right place in DNA replication forks, complex formation with other components in the DNA replication machinery provides an important regulation for UDG function. In this study, we provide the mechanism for EBV UDG BKRF3 nuclear targeting and the interacting domains of BKRF3 with viral DNA replication proteins. Through knockout and complementation approaches, we further demonstrate that in addition to UDG activity, the interaction of BKRF3 with viral proteins in the replication compartment is crucial for efficient viral DNA replication.

In this work, we demonstrate that carbon dots (CDs) can be used as a dispersing agent for graphene as well as a reducing agent for KMnO4 for the synthesis of manganese oxide (MnO x )–graphene hybrid nanocomposites for supercapacitor applications. CDs obtained from the pyrolysis of ammonium citrate under dry heating possess excellent solubility in water due to their oxygen- and nitrogen-containing functional groups. In addition, the sp2-carbon-rich CDs exhibited strong interaction with graphene through π–π stacking for self-immobilizing on graphene in the preparation of water-soluble CD/graphene nanocomposites (CDGs). Interestingly, MnO x could be grown in situ on CDGs after reaction with KMnO4 in aqueous solution under a mild reaction temperature (75 °C). Under the mild reaction conditions, CDs undergo sacrificial oxidation for the formation of MnO x nanoparticles on graphene, whereas the graphene’s graphitic carbons are protected. The as-formed nanostructured MnO x on CDGs (MnO x –CDGs) was employed to fabricate flexible solid-state supercapacitor which exhibited good capacitance properties (specific capacitance ∼280 F g–1) with very high charge–discharge cyclic stability (>10 000 cycles) and good capacitance retention at 90° bending angle. Compared to other graphene-based nanocomposites, our one-pot synthesis route for MnO x –CDGs is relatively green, simple, rapid, and cost-effective and has a great potential for the synthesis of different metal oxide-decorated graphene nanocomposites for energy conversion and storage applications

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The Pediococcus pentosaceus ACCEL bacteriocin was purified to electrophoretical homogeneity by cell adsorption-desorption and Superose 12 fast performance liquid chromatography (FPLC). The purified bacteriocin, with a molecular mass of 17.5 kDa and an N-terminal sequence of -KYYGNGVTXGKHSXXVDXG-, belongs to class IIa and is designated pediocin ACCEL. It was inactivated by various proteases and stable at pH 2.0-6.0 and <100 degrees C. More than 80% activity was left even after 15 min of heating at 121 degrees C and pH 2.0-4.0. Gram-positive food-borne pathogens were inhibited by this bacteriocin, but Gram-negative ones were not. According to the storage stability study, the purified pediocin was stable at pH <6.0 and low temperature. No significant change in bactericidal activity was observed after freeze-drying and subsequent 1-month storage at room temperature.

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