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

Bin Chen, Yian Wang, Ming You

Research output: Contribution to journalArticlepeer-review

Abstract

A tandem repeat region in the second intron of the K-ras gene has been reported to be a possible regulatory site for transcription [You et al. Proc Natl Acad Sci USA. 1992; 89: 5804-5808; Chen et al. Proc Natl Acad Sci USA. 1994; 91: 1589-1593; Chen et al. Carcinogenesis. 1994; 15: 2031-2035]. 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 KS strains (which have K-ras alleles identical to those of the 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 Kr strains (which have K-ras alleles identical to those of the resistant C3H/HeJ strain) and the Ks or Ki strains. Southwestern blot analysis of DNA-protein complexes indicated that the 2 repeated regions might bind the same regulatory complex.

Original languageEnglish (US)
Pages (from-to)179-192
Number of pages14
JournalExperimental Lung Research
Volume31
Issue number2
DOIs
StatePublished - Mar 2005

Keywords

  • K-ras
  • Protein binding sites
  • Second intron

ASJC Scopus subject areas

  • Molecular Biology
  • Pulmonary and Respiratory Medicine
  • Clinical Biochemistry

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