Skip to main content
. 2016 Apr 25;6:24979. doi: 10.1038/srep24979

Figure 1. Processing of inosine by EndoV enzymes.

Figure 1

(a) Schematic illustration of EndoV cleavage of the second phosphodiester bond 3′ to an inosine residue. (b) Increasing amounts of human EndoV (hEV;1–25 nM) and mouse EndoV (mEV; 10–100 nM) were incubated with 1 nM RNA or DNA substrates: ss/ds I-RNA, ss/ds ctr-RNA, ss/ds IIUI-RNA, ss/ds dIrG-DNA and (c) ss/ds I-DNA at 37 °C for 30 minutes using standard reaction buffer. 100 nM of the site specific mEndoV mutant D52A was included in all assays. Cleavage products were analyzed by 20% denaturing PAGE and visualized by phosphorimaging. (d) EndoV cleavage products for the IIUI-RNA substrates were run on 20% sequencing gels alongside with 10, 13 and 15 mer RNA markers. The illustration shows potential EndoV cleavage sites on the IIUI-RNA substrate. Abbreviations: - = no enzyme added, S = substrate, P = cleaved product, ss = single-stranded, ds = double-stranded, EcEV = E. coli EndoV. Enzyme amounts are shown in nM and sizes of RNA/DNA substrates and products are indicated in parentheses.