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Nucleic Acids Research, 1993, Vol. 21, No. 18 4296-4304
© 1993

MOLECULAR BIOLOGY

DNA replication facilitates the action of transcriptional enhancers in transient expression assays

Angus C. wilson+ and Roge K. Patient*

Developmental biology Research Centre, The Randall Institute, Biomedical Sciences Division, King's College London 26-29 Drury Lane, London Wc2B 5RL, UK

*To whom correspondence should be addressed

Received May 27, 1993. Revised August 10, 1993. Accepted August 10, 1993.

We demonstrate a general role for DNA replication in the activation of gene transcription in transient transfectlon assays. The effect is observed for a wide range of genes and cell types, transfected by a number of protocols and is independent of increased template copy number. Replication does not stimulate transcription driven by proximal promoter elements alone but requires a functional enhancer element. This synergy between an active replication origin and an enhancer Is not confined to elements from viruses such as SV40, which undergo an early to late switch In gene expression that Is tightly coupled to replication, since the enhancer-containing long terminal repeats from retrovlruses are strongly stimulated by replication. Furthermore, synthetic enhancers consisting of multlmerised binding sites for one or two factors are also subject to replication-activation. The diversity of synthetic and natural enhancers used In this study suggests that replication and transcription do not share a common protein factor. We propose that replication leads to chromatln modifications that facilitate enhancer action

+Present address: cold spring Harbor Laboratory, Cold Spring Harobor, NY 11724, USA


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