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. 2019 May 28;10(6):411. doi: 10.3390/genes10060411

Figure 5.

Figure 5

The NTD of Pif1 affects DNA binding and helicase activities. (A) Pif1ΔN binds the Tel15 single-stranded DNA (ssDNA) substrate much more tightly than full-length Pif1. The plotted values represent the results of EMSAs using radiolabeled Tel15 and the indicated concentrations of Pif1 and Pif1ΔN protein. (B) Pif1 helicase activity is stimulated by RNA–DNA hybrids. Unwinding of a radiolabeled DNA–DNA fork substrate and an analogous RNA–DNA fork were monitored with native gels at the indicated concentrations of Pif1. (C) Pif1ΔN displays minor helicase activity stimulation in the presence of the RNA–DNA hybrid fork. (D) Both Pif1 and Pif1ΔN have higher affinity for the RNA–DNA hybrid fork substrate the than Tel15 ssDNA. All data points represent the average of ≥3 independent experiments, and the error bars correspond to the standard deviation. The plotted data were fit with hyperbolic curves using GraphPad Prism software.