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. Author manuscript; available in PMC: 2014 Jul 25.
Published in final edited form as: Immunity. 2013 Jul 18;39(1):123–135. doi: 10.1016/j.immuni.2013.07.001

Figure 4. DHX33 requires the helicase C domain for binding to dsRNA.

Figure 4

(A and B) HA-DHX33 was overexpressed in HEK293T cells and purified by anti-HA beads. Purified HA-DHX33 protein was incubated with biotinylated poly I:C (A) or biotinylated genomic reoviral RNA (B) in the absence (0 μg) or presence (A: 5 or 50 μg of poly I:C; B: 50 μg of poly U, 5 or 50 μg of poly I:C) of unlabeled competitor. Biotinylated poly I:C (A) or reoviral RNA (B) was precipitated by streptavidin beads, followed by immunoblotting (IB) for HA-DHX33.

(C and D) Recombinant His-DHX33, generated in E. coli, was incubated with biotinylated poly I:C (C) or biotinylated reoviral genomic RNA (D) in the absence (0 μg) or presence (5 or 50 μg) of unlabeled poly I:C as a competitor. Biotinylated dsRNA was immunoprecipitated by streptavidin beads, followed by IB for DHX33.

(E) Purified full-length and truncated HA-DHX33 proteins were incubated individually with biotinylated poly I:C. Biotinylated poly I:C was immunoprecipitated by streptavidin beads, followed by IB for HA-DHX33. DEAD, Asp-Glu-Ala-Asp domain; HELICc, helicase C-terminal domain; HA2, helicase-associated domain 2; DUF, domain of unknown function.

See also Figure S3.