Kaposi's Sarcoma-Associated Herpesvirus ORF57 Protein Binds and Protects a Nuclear Noncoding RNA from Cellular RNA Decay Pathways
Figure 3
PAN RNA is posttranscriptionally up-regulated by ORF57.
(A) Northern blots showing a dose-dependent response of PAN RNA driven by the CMVIE, EF1α, and SV40 promoters. Relative values and standard deviations are shown below; p values are given for a comparison of the 0 and 0.4 µg ORF57 quantities (CMVIE, n = 3; EF1a and SV40, n = 5). (B) Pol II ChIP results using TRP-Δ79. ChIP assays were done in the presence of dox or in its absence in cells transfected with or without ORF57 as indicated. PCR amplicons were nt 93-147 (5′) and 777-863 (3′) relative to the transcription start site. The diagram shown below is not to scale. Quantitation and background correction are described in the Materials and Methods section (PAN 5′: +dox, n = 4, +/−ORF57, n = 5; PAN 3′: +dox n = 2, +/− ORF57 n = 3). P-values are shown for the +/−ORF57 data sets. (C) Pol II ChIP results with PAN-Δ79; details are the same as (B). (PAN 5′: n = 4; PAN 3′: n = 2). The increased signal for the 3′ samples in the –ORF50 control is a result of lower signal for the 3′ amplicon in the +ORF50 samples and is not due to an increase in the amount of background signal (data not shown).