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. 2004 Jul;24(14):6514–6524. doi: 10.1128/MCB.24.14.6514-6524.2004

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Copyright © 2004, American Society for Microbiology

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FIG. 4. — Binding of Mcm1 to ARS120 and ARS131a. (A) Binding of Mcm1 to fragments I, II (domain C), and III (domains A and B) of ARS120 and ARS131a by EMSA. Endpoints of fragments are indicated on the bar diagram. Fifty femtomoles of DNA was added in each lane. Amounts of Mcm1 added: lanes 1, 5, and 9, 0 pmol; lanes 2, 6, and 10, 2.2 pmol; lanes 3, 7, and 11, 6.7 pmol; lanes 4, 8, and 12, 20 pmol. (B) Competition for binding of Mcm1 by fragment I of ARS120 and ARS131a. Labeled fragment I of ARS120 bound to Mcm1 (4 pmol) was competed with increasing concentrations of unlabeled competitor DNA. (C) DNase protection in the nonconserved region of fragment I by Mcm1. (i) Top strand of ARS131a and ARS120. (ii) Bottom strand of ARS120. Amounts of Mcm1 added: lanes 5, 6, and 11, 0 pmol; lanes 2, 7, and 12, 5 pmol; lanes 3, 8, and 13, 10 pmol; lanes 4, 9, and 14, 20 pmol; lanes 5 and 10, 40 pmol. A bar indicates protection. An asterisk indicates hypersensitivity. Ia, Ib, Ic, and Id are footprints of Mcm1. (D) Sequence of DNase footprints of Mcm1 on fragment I. The protected sequence is underscored, and hypersensitivity is indicated by an asterisk. Protection patterns are deduced from at least two independent gels for both the top and bottom strands of ARS120 and ARS131a.

FIG. 4. — Binding of Mcm1 to ARS120 and ARS131a. (A) Binding of Mcm1 to fragments I, II (domain C), and III (domains A and B) of ARS120 and ARS131a by EMSA. Endpoints of fragments are indicated on the bar diagram. Fifty femtomoles of DNA was added in each lane. Amounts of Mcm1 added: lanes 1, 5, and 9, 0 pmol; lanes 2, 6, and 10, 2.2 pmol; lanes 3, 7, and 11, 6.7 pmol; lanes 4, 8, and 12, 20 pmol. (B) Competition for binding of Mcm1 by fragment I of ARS120 and ARS131a. Labeled fragment I of ARS120 bound to Mcm1 (4 pmol) was competed with increasing concentrations of unlabeled competitor DNA. (C) DNase protection in the nonconserved region of fragment I by Mcm1. (i) Top strand of ARS131a and ARS120. (ii) Bottom strand of ARS120. Amounts of Mcm1 added: lanes 5, 6, and 11, 0 pmol; lanes 2, 7, and 12, 5 pmol; lanes 3, 8, and 13, 10 pmol; lanes 4, 9, and 14, 20 pmol; lanes 5 and 10, 40 pmol. A bar indicates protection. An asterisk indicates hypersensitivity. Ia, Ib, Ic, and Id are footprints of Mcm1. (D) Sequence of DNase footprints of Mcm1 on fragment I. The protected sequence is underscored, and hypersensitivity is indicated by an asterisk. Protection patterns are deduced from at least two independent gels for both the top and bottom strands of ARS120 and ARS131a.