id	author	title	date	pages	extension	mime	words	sentences	flesch	summary	cache	txt
cord-005138-u2lwgyyf	Kou, Yi-Hen	Hepatitis C virus NS4A inhibits cap-dependent and the viral IRES-mediated translation through interacting with eukaryotic elongation factor 1A	2006-08-23		.txt	text/plain	6110	363	51	title: Hepatitis C virus NS4A inhibits cap-dependent and the viral IRES-mediated translation through interacting with eukaryotic elongation factor 1A In this study, we have demonstrated that NS4A specifically interacted with eukaryotic elongation factor 1A (eEF1A) and inhibited both cap-and HCV IRES-dependent protein synthesis. To perform translation inhibition assay in culture cells, plasmids encoding V5-tagged HCV nonstructure proteins NS3, NS4A, NS4B, and NS4A mutants were independently cotransfected with the cap-dependent monocistronic luciferase reporter pCMV-Luc or the bicistronic luciferase reporter pJSS12 into Huh7 cells. To perform cap-dependent in-vitro-translation inhibition assay, GST and GST-NS4A fusion proteins that were recovered from the Sepharose 4B beads as described earlier were preincubated with 5 ll of rabbit reticulocyte lysate (RRL, Promega) at 4°C for 1 h. The results demonstrated that both NS4A and NS4B inhibited HCV IRES-mediated translation to levels similar to those of the cap-dependent translation, whereas no effect was detected with the viral NS3 protein (Figure 2d ).	./cache/cord-005138-u2lwgyyf.txt	./txt/cord-005138-u2lwgyyf.txt