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. 2018 Mar 30;46(12):6304–6317. doi: 10.1093/nar/gky231

Figure 1.

Figure 1.

DDX49 is an ATP dependent RNA helicase. (A) Coomassie stained gel of purified GST tagged DDX39B and GST tagged DDX49. The arrow head indicates the full-length proteins. The predicted molecular size of GST-tagged DDX39B is 76.1 kDa and the GST-tagged DDX49 is 81.2 kDa. (B) The ATPase activity of the purified DDX49 and DDX39B with or without total RNA (left panel) or R13 RNA oligo (right panel). The ATP hydrolysis was assessed with the malachite green-molybdate reagent. Data are represented as mean of three independent experiments, with error bars representing standard deviations. Statistical significance was assessed using a two-tailed Welch's unequal variance t-test. n.s stands for not significant, **P< 0.01 and ***P< 0.001. (C) RNA helicase activity of DDX49 in the presence and absence of ATP. (D) RNA helicase activity of DDX49 and DDX39B in the presence and absence of R13 RNA oligo. RNA helicase activity of (E) DDX49 and (F) DDX39B, in the absence of R13 oligo is dependent on the concentration of respective proteins. The helicase assays were repeated at least twice to confirm the reproducibility. dsRNA stands for duplex RNA oligo prepared from labeled and unlabeled RNA oligos; ssRNA for single stranded labeled RNA oligo. dsRNA was incubated at 100°C for 10 min (Denatured dsRNA) prior to loading onto gel.