Purification of intercalator-released p67, a polypeptide that interacts specifically with the c-fos serum response element.
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.
Schröter
H
H
Zentrum für Molekulare Biologie (ZMBH), Universität Heidelberg, FRG.
Shaw
P E
PE
Nordheim
A
A
D001665:Binding Sites; D002738:Chloroquine / Q000494:pharmacology*; D004268:DNA-Binding Proteins / Q000302:isolation & purification*; D006367:HeLa Cells; D008970:Molecular Weight; D011401:Promoter Regions, Genetic; D011485:Protein Binding / Q000187:drug effects; D011518:Proto-Oncogene Proteins / Q000235:genetics*; D012045:Regulatory Sequences, Nucleic Acid
D004268:DNA-Binding Proteins; D011518:Proto-Oncogene Proteins; D002738:Chloroquine