BLM-interacting region of FEN-1 is required for BLM stimulation of FEN-1 endonucleolytic cleavage of 5′ flap substrate

<p><b>Copyright information:</b></p><p>Taken from "The interaction site of Flap Endonuclease-1 with WRN helicase suggests a coordination of WRN and PCNA"</p><p>Nucleic Acids Research 2005;33(21):6769-6781.</p><p>Published online 2 Dec 2005</p><p>PMCID:PMC1301591.</p><p>© The Author 2005. Published by Oxford University Press. All rights reserved</p> Reaction mixtures (20 µl) containing 10 fmol of the 26 nt 5′ flap substrate, the specified concentrations of wild-type FEN-1, FEN-1ΔC or FEN-1ΔP, and BLM were incubated at 37°C for 15 min under standard conditions as described in Materials and Methods. Products were resolved on 20% polyacrylamide denaturing gels. () Phosphorimage of a typical gel is shown. Lane 1, no enzyme; lanes 2, 4 and 6 are wild-type FEN-1, FEN-1ΔP and FEN-1ΔC, respectively; lanes 3, 5 and 7 are wild-type FEN-1, FEN-1ΔP and FEN-1ΔC, respectively, in the presence of BLM; lane 8, BLM alone. () Per cent incision (mean value of at least three independent experiments with SD indicated by error bars). Quantitative data are shown for incision reactions with FEN-1 or FEN-1ΔC alone (open bars) or in the presence of BLM (light gray bars)