Transcription factor LSF binds two variant bipartite sites within the SV40 late promoter.

The HeLa transcription factor LSF has been purified by heparin-agarose and DNA affinity chromatography, and its DNA binding and transcription properties have been characterized. LSF is a 63-kD polypeptide that binds to two distinct bipartite sites within the SV40 promoter region. One binding site consists of GC motifs 2 and 3 within the 21-bp repeats (LSF-GC site), and the other consists of sequences centered 44 bp upstream of the major late initiation site, L325 (LSF-280 site). Four guanine residues within the LSF-GC site, when methylated, strongly interfere with LSF binding. Alteration of the spacing, but not the sequence, between the two directly repeated GC motifs dramatically reduces the binding affinity of LSF for the site. Thus, LSF appears to recognize directly repeated GC motifs, when their center-to-center distance is 10 bp. The LSF-GC and LSF-280 sites share limited sequence homology. Only half of the LSF-280 site contains a short GC-rich sequence homologous to the GC motif. However, the binding affinity of LSF to the two sites is similar. LSF activates transcription from the SV40 late promoter in vitro from initiation site L325, via its binding to the template DNA.

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