Supplementary Figure 5: The fate of 5'-adenylated BER intermediate in the case of APTX and/or FEN1 deficiency.
From: Role of polymerase β in complementing aprataxin deficiency during abasic-site base excision repair
(a) The dRP lyase, DNA adenylation and FEN1 excision assays with purified enzymes pol β, FEN1, and APTX. Lane 1 is a minus enzyme control, lane 2, 3, and 4 are reference reactions containing FEN1, pol β, and APTX, respectively. The migration position of the DNA substrate (17dRP+AMP, Substrate 2 in Supplementary Table 1) is indicated. (b-g) The dRP lyase, DNA adenylation and FEN1 excision assays in the cell extracts prepared from the yeast strains (Supplementary Table 3). DNA substrate was not pretreated with UDG. The reaction products observed in the cell extracts from the wild-type (b, lanes 5-7), hnt3Δ (c, lanes 8-10), rad27Δ (d, lanes 11-13), hnt3Δrad27Δ (e, lanes 14-16), rad27Δ::POLB (f, lanes 17-19), hnt3Δrad27Δ::POLB (g, lanes 20-22) yeast strains are indicated. In panels a-g, the positions of the products for FEN1 excision cleavage (16- or 15-mer), pol β lyase (17-mer), and APTX removal of AMP (17dRP) are indicated. Three lanes in each set (in panels b-g) correspond to the time points 5, 15, or 30 min.
