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.
Chen
Bin
B
Department of Pathology, Medical College of Ohio, Toledo, Ohio, USA.
Wang
Yian
Y
You
Ming
M
D000225:Adenine; D000818:Animals; D001483:Base Sequence; D001665:Binding Sites; D026501:Blotting, Southwestern; D002460:Cell Line; D018983:DNA Footprinting; D003850:Deoxyribonuclease I; D004591:Electrophoresis, Polyacrylamide Gel; D011905:Genes, ras; D014644:Genetic Variation; D007438:Introns; D051379:Mice; D008815:Mice, Inbred Strains; D008969:Molecular Sequence Data; D009687:Nuclear Proteins / Q000378:metabolism*; D020080:Tandem Repeat Sequences; D013941:Thymine
D009687:Nuclear Proteins; D003850:Deoxyribonuclease I; D000225:Adenine; D013941:Thymine