TABLE 4.
Summary of relative affinities (Krel) and off-rates (T1/2) for the HPV E2sct proteins with DNA sites
| Clone | Sequence | Krela | T1/2 (min)b |
|---|---|---|---|
| BS3-wt | cagaACCGTTTTGGGTtccc | 0.09 | >90 |
| BS3-t→a | cagaACCGATTTGGGTtccc | 0.11 | >90 |
| BS3-g→t | cagaACCGTTTTGTGTtccc | 0.13 | >90 |
| BS1-wt | cagaACCGAAACCGGTtccc | 0.22 | >90 |
| BS2-wt/BS1-c→t | cagaACCGAAATCGGTtccc | 1.00 | >90 |
| hE2BS | cagaACCGATTTGTGTtccc | 0.24 | >90 |
| hFLIP | cagaACACAAATCGGTtccc | 0.78 | >90 |
| hE2BS-KO | cagaGTACAAATCGACtccc | <0.01 | N.D. |
Krels were calculated based on the amount of unlabeled competitor DNA (Clone column) necessary to compete away 10,000 cpm (∼1 nM) of radiolabeled BS2-wt probe in at least three experiments.
T1/2 is equal to the amount of time (minutes) that a 1,000-fold excess of unlabeled BS2-wt competitor DNA requires to compete away 50% of a 10,000 cpm (∼1 nM) radiolabeled clone probe (Clone column) in at least three experiments. N.D., not determined. Within the sequences, lowercase lettering corresponds to flanking nucleotides, uppercase letters correspond to the 12-bp E2BS, and the underlined letters correspond to those nucleotides which differ from the wild type for each site.