Figure 4.

MHV ns2 Protein Inhibits Ribosomal RNA Degradation as well as Accumulation of 2-5A in Infected BMMs and in 293T Cells Induced with Poly(I:C)

(A) B6 BMMs were infected (1 PFU/cell). At the indicated times, RNA was purified and analyzed on a Bioanalyzer. The positions of 18S and 28S rRNA are indicated.

(B) 293T cells were transfected with pCAGGS (pC), pC-ns2, or pC-ns2-H126R and 24 hr later were transfected with 10 μg/ml poly(I:C). Four hours later, RNA was purified and analyzed on a Bioanalyzer.

(C and D) 2-5A was isolated from virus-infected BMMs described in (A) derived from B6 mice at 6 and 9 hr p.i (C) or RNase L^−/− mice at 9 hr p.i. (D) and analyzed by FRET. Error bars represent the SEM (n = 3). Asterisks indicate that differences are statistically significant (^∗p < 0.05 and^∗∗∗p < 0.001).

(E) 2-5A from transfected 293T cells described in (B) was isolated and quantified by FRET. Error bars represent the SEM (n = 3). Asterisks indicate that differences are statistically significant (^∗∗∗p < 0.001).

See also Figure S2.