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. 2012 Jan 31;22(6):973–985. doi: 10.1038/cr.2012.18

Figure 6.

Figure 6

Processing enhancement by MutLα requires integrity of the MutLα ATPase and pri-miRNA binding activity. (A) Effect of ATP on miRNA processing stimulation by MutLα. In-vitro processing of pri-miR-422a was carried out in the presence of 2 mM ATP or AMP-PNP, as indicated. pri-miR-422a and the resulting pre-miR-422a and 3′-, 5′-products are indicated with arrows. (B) ATPase assay of wild-type (WT) and mutant MutLα proteins. The indicated proteins were incubated with 330 mM (γ-32P)-ATP in reactions containing 50 mM HepesċKOH, pH7.6 and 10 mM MgCl2, and the hydrolysis products were analyzed in a 20% polyacrylamide gel. (C) In-vitro processing of pri-miR-422a using WT MutLα, MutLαEA (EA), MutLαF99L (F99L) or MutLαE705K (E705K). (D) In-vitro processing of WT pri-miR-16-1 and flanking basal segment deleted pri-miR-16-1ΔBS (ΔBS). *Non-specific reaction products.