Interactions of the high‐mobility‐group‐like Ceratitis capitata C1 proteins with DNA

We have studied the interactions of the high-mobility-group-like proteins (C1a1, C1a2 and C1b) from the fruit fly Ceratitis capitata with DNA. Nitrocellulose filter binding assays, thermal denaturation studies and spectrofluorimetry of the complexes revealed the existence of specific and nonspecific interactions. Thermal denaturation curves showed that the three proteins stabilized the DNA, thus suggesting a preferential binding to double-stranded DNA. The calculation of the thermodynamic parameters of the interactions showed that the nonspecific bindings were characterized by low association constants (Ka) with values ranging from 2.7×104 M−1 to 2.0 × 106 M−1. Also, the cooperativity of these interactions was relatively high (cooperativity factor, w, values ranging over 20–35), and the number of nucleotides involved was low (1–3 base pairs). On the other hand, the existence of specific interactions between C1 proteins and DNA was suggested by two facts: (a) the retention of C. capitata [3H]DNA in nitrocellulose filters was only a low percentage of total input DNA and (b) there was a marked size dependence of the binding (25% retention of a 40-kb DNA and only 3% retention with a DNA of 1 kb). The specific bindings had higher Ka values than the nonspecific ones, and they also were cooperative. Some differences were observed between C1b and the C1a proteins about the way they interact with C. capitata DNA.