ETHIDIUM-BROMIDE PROVIDES A SIMPLE TOOL FOR IDENTIFYING GENUINE DNA-INDEPENDENT PROTEIN ASSOCIATIONS

被引：429

作者：

LAI, JS ^{[1
]}

HERR, W ^{[1
]}

机构：

[1] SUNY STONY BROOK,GENET PROGRAM,STONY BROOK,NY 11794

来源：

PROCEEDINGS OF THE NATIONAL ACADEMY OF SCIENCES OF THE UNITED STATES OF AMERICA | 1992年 / 89卷 / 15期

关键词：

POU PROTEINS; HUMAN AUTOANTIGEN KU; IMMUNOPRECIPITATION; PROTEIN-AFFINITY CHROMATOGRAPHY; CONTAMINATING DNA;

D O I：

10.1073/pnas.89.15.6958

中图分类号：

O [数理科学和化学]; P [天文学、地球科学]; Q [生物科学]; N [自然科学总论];

学科分类号：

07 ; 0710 ; 09 ;

摘要：

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.

引用

页码：6958 / 6962

页数：5

共 41 条

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