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. 1999 Jun 8;96(12):6694–6699. doi: 10.1073/pnas.96.12.6694

Figure 5.

Figure 5

RNA oligonucleotide-competition experiments confirm a GRSF-1-positive translational regulatory function. (A and B) RNA oligonucleotide-competition experiments. Template cellular 5′ UTR SEAP-LUC (A) or viral 5′ UTR NP-LUC (B) was translated in an influenza virus-infected HeLa cell extract in the absence or presence of various competitors as indicated below each bar. Competitor RNAs were added at 200 × molar ratio to template. The nucleotide sequences of competitors are shown in Fig. 2A. An arbitrary value of 100 represented 4,215 cpm SEAP-LUC of luciferase activity per μl of HeLa cell extract in A and 4,520 cpm of NP-LUC luciferase activity per μl of HeLa cell extract in B. (C) Effects of GRSF-1 reconstitution in RNA oligonucleotide-compromised extracts. Either template NP-LUC or SEAP-LUC was translated in a virus-infected HeLa extract in the absence (No comp) or presence of competitor NP 5′ UTRs (NP) or in the presence of competitor NP 5′ UTRs plus 0.200 μg of GST-GRSF-1 (NP+GRSF-1) as indicated below each bar. After 45 min at 30°C, translation products were assayed by using a Luciferase Assay System. A value of 100 represented 4,240 cpm of NP-LUC luciferase activity per μl of HeLa cell extract in C. Other values then were determined relative to this standard. Values throughout are the mean ± SD of three experiments per group.