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. 1998 Sep;72(9):7484–7493. doi: 10.1128/jvi.72.9.7484-7493.1998

FIG. 2.

FIG. 2

PPBP binds through the minor groove of DNA. (A) Formation of polh B domain-PPBP complex is inhibited by actinomycin D. DNA-protein complex obtained with 1 μg of Sf9 cell nuclear extract (lane 9) was incubated with 10 (lane 3), 25 (lane 4), 50 (lane 5), 125 (lane 6), 250 (lane 7), or 500 (lane 8) μM actinomycin D. As controls, labeled polh B domain was incubated either alone (lane 1) or with 500 μM actinomycin D (lane 2). polh B domain spans from −63 to −32 on the p29 promoter, as shown in Fig. 1. (B) Formation of polh B domain-PPBP complex is inhibited by distamycin A. DNA-protein complex obtained with 1 μg of Sf9 cell nuclear extract (lane 3) was incubated with 10 (lane 4), 25 (lane 5), 50 (lane 6), 100 (lane 7), 200 (lane 8), 500 (lane 9), or 1,000 (lane 10) μM distamycin A. As controls, labeled polh B domain was incubated either alone (lane 1) or with 1,000 μM distamycin A (lane 2). (C) Formation of polh B domain-PPBP complex is inhibited by Hoechst 33258 dye. DNA-protein complex obtained with 1 μg of Sf9 cell nuclear extract (lane 3) was incubated with 10 (lane 4), 25 (lane 5), 50 (lane 6), 100 (lane 7), 200 (lane 8), 500 (lane 9), or 1,000 (lane 10) μM Hoechst 33258. As controls, labeled polh B domain was incubated either alone (lane 1) or with 1,000 μM Hoechst 33258 (lane 2). (D) The polh B domain-PPBP complex is unaffected by increasing concentrations of methyl green. Labeled polh B domain was incubated either alone (lane 1), with 5 mM methyl green (lane 2), or with 1 μg of Sf9 cell nuclear extract (lanes 3 to 11). The DNA-protein complex obtained (lane 3) was incubated with 25 (lane 4), 50 (lane 5), 125 (lane 6), 250 (lane 7), 500 (lane 8), 1.25 (lane 9), 2.5 (lane 10), or 5 (lane 11) mM methyl green.