Identification and purification of a polypeptide that binds to the c-fos serum response element

A short DNA sequence element, the serum response element (SRE), which binds a nuclear protein, serum response factor (SRF), mediates transient transcriptional activation of c-fos and cytoskeletal actin genes in response to serum factors. Variant SRE sequences with different affinities for HeLa cell SRF were synthesised. Binding of SRF to these sites in vitro correlates with the transcriptional properties of these elements in vivo, suggesting that SRF is a positively acting transcription factor. A 67-kd polypeptide was identified as the DNA-binding component of SRF by photoactivated DNA-protein cross-linking in vitro. The high affinity SRF-binding site was used to purify this polypeptide to virtual homogeneity in a single DNA affinity chromatography step.