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. 1988 Feb;8(2):982–987. doi: 10.1128/mcb.8.2.982

Capturing nuclear sequence-specific DNA-binding proteins by using simian virus 40-derived minichromosomes.

U Nir 1, E Fodor 1, W J Rutter 1
PMCID: PMC363232  PMID: 2832746

Abstract

We have used recombinant simian virus 40 (SV40) minichromosomes to retrieve sequence-specific DNA-binding proteins derived from the cell nucleus of COS-7 cells. We showed that the transcription factors AP-1 and Sp1 are stably bound to the SV40 DNA late in viral infection. Under similar conditions, minichromosomes carrying the rat insulin (rINS1) enhancer, which is under negative regulation in COS-7 cells, bound two proteins which mapped to distinct regions of the rINS1 enhancer. The SV40 P element competed for one of these proteins which bound to the region from -198 to -230. This factor may be related to AP-1. The other factor selectively bound a regulatory element in the region from -92 to -124 of the insulin enhancer. These proteins may play a role in regulating the rINS1 enhancer function.

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