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. 2019 Feb 14;47(7):3699–3710. doi: 10.1093/nar/gkz057

Figure 3.

Figure 3.

Extraneous RNA inhibited the displacement reaction of Ded1 by sequestering the protein in a nonproductive state. Single-stranded (ssRNA) or double-stranded (dsRNA) RNA was added in increasing amounts to reactions containing 10 nM Ded1, 1 mM ATP and 200 nM of 5′ss-11 bp RNA, which was complementary to the DNA. The 5′ss-11 bp RNA was used at large excess over the fixed DNA, and consequently it also represented an ssRNA competitor for binding Ded1. The ssRNA (RNA01) was a 25-nucleotide-long oligonucleotide with little or no complementarity to the DNA. The dsRNA was made by hybridizing RNA01 to its complementary strand (RNA02). The Fhold was at 7 pN. Data were fit to an exponential equation, P(RNA) = a + b*exp(Kinhibit*[RNA]), where a and b are constants and Kinhibit is the affinity for the competitors. The values of |Toff| are shown with the standard errors of the regression.