TABLE 1.

Inferred differences in included DNA bend angles (Δθ)

Values shown represent differences between bend angles of native and variant complexes as estimated at 4 °C as described previously (39, 40). Absolute bend angles as inferred from PGE can depend on gel composition. The native bend angle using DNA site 5′-CGCATTGTTATCA-3′ (and complement) is thus estimated to be 80° from PGE data obtained in 10% (29:1) gels and 72° from data obtained in 8% (29:1) gels. Despite such variation, the precision of PGE-based estimates of DNA bend angles under a given set of conditions is in general ±1°, implying uncertainties in Δθ values of ±2°. ND, not determined.

Substitution	5′-ATTGTT-3′	5′-TTTGTG-3′a	5′-ATTGAT-3′b
Wild type		−5°	−16°
V5L	<0.5°c	−3°	−20°
V5A	−0.5°c	−7°	ND

^a The native bend angle using the DNA site 5′-GGTGTTTGTGCAGG-3′ (and complement) is estimated to be 74° from data obtained in 10% (58:1) gels (see supplemental Fig. S15). A related site was employed in the solution structure of the SRY-DNA complex (28).

^b The T→A transversion, which adjoins the minor wing of SRY near Val-60, leads to a marked loss of SRY binding affinity (31).

^c No qualitative differences were observed by inspection of PGE patterns as defined side-by-side in the same gel (see Fig. 5, C and D); such relative patterns (i.e. whether a mutation leads to increased or decreased DNA bending) are robust to gel composition.