Lai, J. S., Herr, W. (August 1992) Ethidium bromide provides a simple tool for identifying genuine DNA-independent protein associations. Proc Natl Acad Sci U S A, 89 (15). pp. 6958-62. ISSN 0027-8424 (Print)0027-8424 (Linking)
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Abstract
DNA-dependent and DNA-independent associations of DNA-binding proteins are important in transcriptional regulation. The analysis of DNA-independent associations frequently relies on assaying protein interaction in the absence of target DNA sequences. We have found that contaminating DNA in protein preparations can stabilize DNA-dependent associations that may appear DNA-independent. Three cellular proteins of 70, 85, and 110 kDa coimmunoprecipitated with the octamer motif-binding protein Oct-2 because of the presence of contaminating DNA in the cell extracts. In addition, heterodimer formation between Oct-1 (or Oct-2) and Pit-1 during protein-affinity chromatography was stabilized by the contaminating DNA. In both instances, these DNA-dependent protein associations were selectively inhibited by ethidium bromide in the precipitation reaction without any evident effect on DNA-independent protein associations. Thus, ethidium bromide may serve as a simple and general indicator of DNA-dependent and DNA-independent protein associations.
| Item Type: | Paper |
|---|---|
| Uncontrolled Keywords: | Animals Base Sequence Chromatography, Affinity DNA/genetics/isolation & purification/*metabolism DNA-Binding Proteins/isolation & purification/*metabolism *Ethidium Glutathione Transferase/genetics/isolation & purification/*metabolism Host Cell Factor C1 Immunoglobulin Heavy Chains/genetics Indicators and Reagents Micrococcal Nuclease Molecular Sequence Data Molecular Weight Octamer Transcription Factor-1 Oligodeoxyribonucleotides Promoter Regions, Genetic Protein Binding Recombinant Fusion Proteins/isolation & purification/metabolism Ribonucleases Transcription Factor Pit-1 Transcription Factors/isolation & purification/metabolism Transfection |
| Subjects: | Investigative techniques and equipment bioinformatics > genomics and proteomics > genetics & nucleic acid processing > protein structure, function, modification > protein types > DNA binding protein |
| CSHL Authors: | |
| Communities: | CSHL labs > Herr lab |
| Depositing User: | Matt Covey |
| Date: | 1 August 1992 |
| Date Deposited: | 22 Sep 2015 16:18 |
| Last Modified: | 09 Nov 2017 20:14 |
| PMCID: | PMC49624 |
| Related URLs: | |
| URI: | https://repository.cshl.edu/id/eprint/31855 |
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