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. 2011 Aug 11;39(21):9238–9249. doi: 10.1093/nar/gkr653

Figure 1.

Figure 1.

TWINKLE can efficiently load onto circular DNA substrates. TWINKLE interactions with circular ssDNA, linear ssDNA and linear dsDNA. (A) TWINKLE–DNA interactions were monitored by EMSA using 32P-labeled circular ssDNA (100 nt, upper panel), linear dsDNA (100 bp, middle panel) or linear ssDNA (100 nt, lower panel) as described in ‘Materials and Methods’ section. Incubation temperatures and the amount of TWINKLE (0.1 or 0.2 pmol) are indicated at the top of the figure. Lanes 7–12 and 14–19, were cross-linked with glutaraldehyde and lanes 14–19; were further treated with SDS and heated at +95°C for 5 min. (B) The binding of TWINKLE to circular ssDNA is nucleotide independent. Binding reactions between TWINKLE and closed circular ssDNA in the absence (lanes 1–4) or presence of UTP (lanes 5–8) were performed as described in ‘Materials and Methods’ section. Lanes 1 and 5, DNA substrate alone; lanes 2–4 and 6–8, were incubated with 200 fmol TWINKLE at +4°C; lanes 3–4 and 7–8, were cross-linked with glutaraldehyde; and lanes 4 and 8 were further treated with SDS and heated at +95°C for 5 min.