(A) Map of domains on the Scm3 open reading frame.
(B) Binding of CEN3 DNA to Scm3(25–247), Scm3(1–113), Scm3(113–247), and Scm3(80–211), shown by EMSA and SYBR Green staining. Essentially full-length, His-tagged Scm3(25–247) or Scm3 subfragments (Figure S1) were mixed with CEN3 DNA (protein:DNA ratio rm = 0.26, 0.52, and 1.04) in 2M NaCl and diluted to 0.2M NaCl in a final volume of 50 μl. After incubation at room temperature for 30 min, samples were analyzed by EMSA and SYBR Green staining.
(C and D) Scm3(1–113) has preference for CEN over 601 DNA (see also Figure S3). Binding of a mixture of CEN and 601 DNA (1 μg each) to Scm3(1–113) (protein:DNA ratio rm = 0.52, 1.04, 2.08) was analyzed as in (A). Graph shows percent unbound DNA to Scm3(1–113):DNA ratios.