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

Bin Chen; Yian Wang; Ming You

doi:10.1080/0190214049049552

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

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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 K^S 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 K^r strains (which have K-ras alleles identical to those of the resistant C3H/HeJ strain) and the K^s or Kⁱ strains. Southwestern blot analysis of DNA-protein complexes indicated that the 2 repeated regions might bind the same regulatory complex.

Original language	English (US)
Pages (from-to)	179-192
Number of pages	14
Journal	Experimental Lung Research
Volume	31
Issue number	2
DOIs	https://doi.org/10.1080/0190214049049552
State	Published - 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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10.1080/0190214049049552

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title = "Characterization of two protein-binding sites in the second intron of the mouse K-ras gene",

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.",

keywords = "K-ras, Protein binding sites, Second intron",

author = "Bin Chen and Yian Wang and Ming You",

note = "Funding Information: Received 10 March 2004; accepted 5 April 2004. This work was supported by NIH Grant R01CA58554. The authors thank Dr. Frederick L. Tyson for the CMT64 and SPON4 cell lines, Dr. Alvin Malkinson for the C-10 cell line. The authors are indebted to Kathy Deanda and Cynthia Kaye for their help in the preparation of this work. Address correspondence to Ming You, Department of Surgery, Washington University School of Medicine, 660 South Euclid, Campus Box 8109, St Louis, MO 63110, USA. E-mail: youm@msnotes. wustl.edu Copyright: Copyright 2008 Elsevier B.V., All rights reserved.",

year = "2005",

month = mar,

doi = "10.1080/0190214049049552",

language = "English (US)",

volume = "31",

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journal = "Experimental Lung Research",

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AU - Chen, Bin

AU - Wang, Yian

AU - You, Ming

N1 - Funding Information: Received 10 March 2004; accepted 5 April 2004. This work was supported by NIH Grant R01CA58554. The authors thank Dr. Frederick L. Tyson for the CMT64 and SPON4 cell lines, Dr. Alvin Malkinson for the C-10 cell line. The authors are indebted to Kathy Deanda and Cynthia Kaye for their help in the preparation of this work. Address correspondence to Ming You, Department of Surgery, Washington University School of Medicine, 660 South Euclid, Campus Box 8109, St Louis, MO 63110, USA. E-mail: youm@msnotes. wustl.edu Copyright: Copyright 2008 Elsevier B.V., All rights reserved.

PY - 2005/3

Y1 - 2005/3

N2 - 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.

AB - 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.

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

Abstract

Keywords

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