Characterization of two protein-binding sites in the second intron of the mouse K-ras gene

A tandem repeat region in the second intron of the K-ras gene has been reported to be a possible regulatory site for transcription. In this study, a second protein-binding site was identified and characterized. It lies downstream (nucleotides 463 to 509) of the tandem repeat region. A T--> C base variation at nucleotide 494 was found in all K(S) strains (which have K-ras alleles identical to those of susceptible A/J strain) and all K(i) strains (which have K-ras alleles identical to those of the intermediate CBA/J strain). DNase I footprint analysis indicated a protein binding site within the downstream repeated region in the second intron of the K-ras gene. Gel mobility-shift studies showed differential protein-binding patterns between the K(r) strains (which have K-ras alleles identical to those of the resistant C3H/HeJ strain) and the K(s) or K(i) strains. Southwestern blot analysis of DNA-protein complexes indicated that the 2 repeated regions might bind the same regulatory complex.