PMC:1140370 / 10978-12745 JSONTXT

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With the Bub66/T-rich substrate, strong protection was detected on the entire single-stranded regions of both top and bottom strands with increasing concentration of Mcm4/6/7, consistent with high affinity of Mcm4/6/7 to T-rich bubble sequences (Figure 1A). Similar, but less significant protection was detected on the single-stranded DNA segment with P1 nucleases (Figure 1A), which is specific to single-stranded DNA. The results indicate that the Mcm complex is loaded onto bubble DNA through binding to both strands of the single-stranded region. Careful examination of Mcm4/6/7 footprints on a bubble indicated strong protection on the 5′-half of the top strand and moderate protection on the remaining 3′-half, as well as on several base-pair duplex segment adjacent to the branch point (Figure 1A and data not shown). A similar pattern of protection was observed also on the bottom strand. 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