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[en] Highlights: ► Depletion of hINO80 significantly reduced PCNA ubiquitination. ► Depletion of hINO80 significantly reduced nuclear dots intensity of RAD18 after UV irradiation. ► Western blot analyses showed phosphorylated hINO80 C-terminus. ► Overexpression of phosphorylation mutant hINO80 reduced PCNA ubiquitination. -- Abstract: Double strand breaks (DSBs) are the most serious type of DNA damage. DSBs can be generated directly by exposure to ionizing radiation or indirectly by replication fork collapse. The DNA damage tolerance pathway, which is conserved from bacteria to humans, prevents this collapse by overcoming replication blockages. The INO80 chromatin remodeling complex plays an important role in the DNA damage response. The yeast INO80 complex participates in the DNA damage tolerance pathway. The mechanisms regulating yINO80 complex are not fully understood, but yeast INO80 complex are necessary for efficient proliferating cell nuclear antigen (PCNA) ubiquitination and for recruitment of Rad18 to replication forks. In contrast, the function of the mammalian INO80 complex in DNA damage tolerance is less clear. Here, we show that human INO80 was necessary for PCNA ubiquitination and recruitment of Rad18 to DNA damage sites. Moreover, the C-terminal region of human INO80 was phosphorylated, and overexpression of a phosphorylation-deficient mutant of human INO80 resulted in decreased ubiquitination of PCNA during DNA replication. These results suggest that the human INO80 complex, like the yeast complex, was involved in the DNA damage tolerance pathway and that phosphorylation of human INO80 was involved in the DNA damage tolerance pathway. These findings provide new insights into the DNA damage tolerance pathway in mammalian cells.
[en] Highlights: •Composite sulfur electrodes are prepared by high-temperature mechanical milling. •The composite exhibited a high discharge capacity of greater than 1200 mAh g−1. •Novel structure unit forms via a reaction between thio-LISICON and sulfur. •Liquid phase sulfur prevents severe decompositions of materials employed. -- Abstract: Composite sulfur electrodes are prepared by high-temperature mechanical milling (443 K) for use in all-solid-state lithium–sulfur batteries, and their structures and electrochemical properties are investigated. Composites comprising sulfur, acetylene black, and a Li3.25Ge0.25P0.75S4 solid electrolyte are fabricated by planetary ball milling using a temperature-controlled system. The composite electrode exhibits a high discharge capacity of greater than 1200 mAh g−1 and a good cycle capability. As a result of high-temperature milling, composites are formed, incorporating novel structural units from the reaction between sulfur and the solid electrolyte, along with their intrinsic characteristics. Hence, high-temperature milling demonstrates promise for the fabrication of a composite electrode exhibiting high, reversible electrochemical activities for use in an all-solid-state lithium–sulfur battery.
[en] Crystals of 6-aminohexanoate-oligomer hydrolase have been obtained by the sitting-drop vapour-diffusion method using sodium citrate as a precipitant. Diffraction data for native and K2PtCl4-derivative crystals were collected to resolutions of 2.00 and 2.20 Å, respectively. 6-Aminohexanoate-oligomer hydrolase (NylC) from Agromyces sp. KY5R was expressed in Escherichia coli JM109 and purified by ammonium sulfate fractionation, anion-exchange column chromatography and gel-filtration chromatography. NylC was crystallized by the sitting-drop vapour-diffusion method with sodium citrate as a precipitant in 0.1 M HEPES buffer pH 7.5 containing 0.2 M NaCl. Diffraction data were collected from native and K2PtCl4-derivative crystals to resolutions of 2.00 and 2.20 Å, respectively. The obtained crystal was plate-shaped, with an I-centred orthorhombic space group and unit-cell parameters a = 155.86, b = 214.45, c = 478.80 Å. The anomalous difference Patterson map of the K2PtCl4-derivative crystal suggested that the space group was I222 rather than I212121