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Next Generation Sequencing of Wild Type and Mice Knocked Out for Known Methionine Sulfoxide Reducatases

ABSTRACT: Methionine sulfoxide reductase (Msr) is an important antioxidant enzyme. The mammalian Msr family has four members: MsrA, MsrB1, MsrB2, and MsrB3. We generated a mouse strain with deletion of all four Msr proteins. Deletion was confirmed by Western blotting. The quadruple knockout mouse is viable, and growth, development, and fertility appear normal. However, they are unexpectedly resistanct to oxidative stresses. We therefore determined the RNA expression profile in liver and heart of wild-type and quadruple knockout mice.

ORGANISM(S): Mus musculus

PROVIDER: GSE130394 | GEO | 2019/10/01

REPOSITORIES: GEO

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Project description:Beech trees produce seeds irregularly, necessitating the storage of these seeds for forestation. Despite the acquisition of desiccation tolerance during development, beech seeds lose viability during long-term storage faster than orthodox-type seeds. Seeds stored for short (3 years) and long (20 years) periods under optimal conditions were compared. Aged seeds characterized with germination capacity reduced to 30% displayed increased membrane damage, manifested as electrolyte leakage and lipid peroxidation levels. Analyses have been based on embryonic axes containing higher levels of reactive oxygen species (ROS) and higher levels of protein-bound methionine sulfoxide (MetO) in aged seeds. Using gel-free shotgun proteomics, 3,949 proteins were identified and 2,442 were reliably quantified to indicate 25 proteins with upregulated abundance and 35 with downregulated abundance in beech seeds under long-term storage compared to those under short-term storage. Functional analyses based on gene ontology annotations revealed that nucleic acid binding activity (biological process), hydrolases (molecular function), ribosome organization or biogenesis and transmembrane transport (cellular processes), and ATP synthesis (pathway) were affected in aged seeds. To verify whether MetO the oxidative posttranslational modification of proteins reversed via the action of methionine sulfoxide reductase (Msr) enzymes is involved in ageing of beech seeds we identified 316 and reliably quantified 226 MetO-containing proteins, among which 9 and 19 exhibited significantly up- and downregulated MetO levels, respectively, in beech seeds under long-term storage. Several Msr isoforms were identified and recognized as MsrA1-like, MsrA4, MsrB5 and MsrB5-like in beech seeds. Only MsrA1-like displayed decreased abundance in aged seeds. We demonstrated that the loss of membrane integrity reflected in elevated abundance of membrane proteins had higher impact on seed ageing progress rather than MetO/Msr system.

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Next Generation Sequencing of Wild Type and Mice Knocked Out for Known Methionine Sulfoxide Reducatases

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