FIG. 2.

(A) ssDNA-binding activity of wild-type MacRPA1 and its chimeras. A fixed amount (1 pmol) of ³²P-labeled ssDNA (lane 1) was incubated with increasing amounts (1, 2, 3, 4, and 5 pmol [indicated by the height of the triangle above the gel) of wild-type MacRPA1 or each of its chimeras (lanes 2 to 6). The MacRPA1 chimera under investigation is indicated above each panel as follows: wild-type MacRPA1 (i), MacRPA1 chimera-1 (ii), MacRPA1 chimera-2 (MbuRPA1-like) (iii), MacRPA1 chimera-3 (iv), MjaRPA-like protein (v), and MthRPA-like protein (vi). The positions of free DNA and protein-DNA complex (protein-bound ssDNA) are shown by the black arrows labeled I and II to the left of the gels, respectively. (B) The ssDNA-binding and dsDNA-binding activities of wild-type MacRPA1 and its chimeras. A fixed amount (1 pmol) of ³²P-labeled ssDNA (lane 1) was incubated with 5 pmol each of MacRPA1 or its derivative (lane 2) and challenged with 1 pmol of unlabeled ssDNA (lane 3), 10 pmol of unlabeled ssDNA (lane 4), 50 pmol of unlabeled ssDNA (lane 5), 1 pmol of unlabeled dsDNA (lane 6), 10 pmol of unlabeled dsDNA (lane 7), and 50 pmol of unlabeled dsDNA (lane 8). The MacRPA1 chimera under investigation is indicated under each panel as follows: wild-type MacRPA1(i), MacRPA1 chimera-1 (ii), MacRPA1 chimera-2 (MbuRPA1-like protein) (iii), MacRPA1 chimera-3 (iv), MjaRPA-like protein (v), and MthRPA-like protein (vi). Free DNA and protein-DNA complex were resolved by 8% polyacrylamide gel electrophoresis followed by visualization using autoradiography. The positions of free DNA and protein-DNA complex (protein-bound ssDNA) are shown by the black arrows labeled I and II to the left of the gels, respectively.