Purification of intercalator-released p67, a polypeptide that interacts specifically with the c-fos serum response element. |
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| Authors: | H Schr?ter P E Shaw A Nordheim |
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| Affiliation: | Zentrum für Molekulare Biologie (ZMBH), Universität Heidelberg, FRG. |
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| Abstract: | Incubation of intact nuclei in buffers containing the DNA intercalating drug chloroquine leads to release of proteins that interact with DNA. We demonstrate here that a protein which binds to a motif within the human c-fos promoter, identified as the serum response element (SRE), is quantitatively released from HeLa nuclei, whereas nuclear factor 1 (NF 1) is not. Purification of the SRE binding protein by affinity chromatography to greater than 95% homogeneity allowed us to identify it as a polypeptide of approximately 67,000 daltons. The DNA contacts made by p67, as identified by methylation interference experiments, are indistinguishable from those of the serum response factor described previously. |
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