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• W2085778157 abstract "Abstract Intracerebral infection with Theiler’s murine encephalomyelitis virus (TMEV) induces a demyelinating disease that resembles human multiple sclerosis. In order to delineate the early events in this virus‐induced neuroinflammatory disease, we have analyzed global GTPases gene activation following TMEV infection of murine brain astrocytes. DNA hybridization microchip analysis demonstrated that 10 sequences described as GTPbinding proteins and GTPases in different protein databases were over‐expressed, in response to this infectious agent in astroglial cells. We have first characterized both the GTP‐binding and GTPase activities in uninfected astrocyte membranes from a biochemical point of view. The increase in such activities was further validated in TMEV‐infected astrocytes, peaking 2–4 h after infection. Over‐expression is also induced by the inflammation‐related chemokines interleukin‐6 and interferon‐γ but not by interleukin‐1α or tumor necrosis factor‐α. From the many GTPases that could be over‐expressed we have studied two, because of its biological significance; Ras p21 and the subunit αi2 of G proteins. Western blots revealed increases in both proteins after infection with TMEV, in accordance with the previous enzymologic results. An increase in the active form of Ras (the GTP bound form) in cell lysates was also confirmed by affinity binding to a glutathione‐S‐transferase‐fusion protein, following TMEV infection. A final demonstration of physiological up‐regulation is provided by UV cross‐linking of membrane proteins with the hydrolysis‐resistant GTP agonist GTP [γ‐ 35 S]. This technique allow us to detect, after SDS‐PAGE, the increase of two further majoritary GTPbinding proteins with MW of 62 and 49 KDa. A quantitative analysis of four selected genes coding for p21 ras, Gαi2 subunit of protein G, Munc‐18 and protein interacting with C kinase 1, was performed by real‐time RT‐PCR to verify the microarray results. The study of GTPase activity and of the above genes by RT‐PCR in brains of sick mice, demonstrated a significative increase in mRNA coding for p21ras and protein interacting with C kinase 1 in vivo . Here we demonstrate that one of the mechanisms triggered by TMEV infection of astrocytes is the up‐regulation of proteins related to GTP metabolism, one important signal transduction system in mammalian cells." @default.
• W2085778157 created "2016-06-24" @default.
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• W2085778157 date "2007-10-08" @default.
• W2085778157 modified "2023-10-17" @default.
• W2085778157 title "Over-expression of GTP-binding proteins and GTPase activity in mouse astrocyte membranes in response to Theiler’s murine encephalomyelitis virus infection" @default.
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• W2085778157 doi "https://doi.org/10.1111/j.1471-4159.2007.05020.x" @default.
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