The polyomavirus enhancer activates chromatin accessibility on integration into the HPRT gene.
Recent studies suggest that enhancers may increase the accessibility of chromatin to transcription factors. To test the effects of a viral enhancer on chromatin accessibility, we have inserted minigenes with or without the polyomavirus enhancer into the third exon of the hypoxanthine phosphoribosyltransferase (HPRT) gene by homologous recombination and have prepared high-resolution maps of gene accessibility by using a novel polymerase chain reaction assay for DNase I sensitivity. In its native state, we find that the HPRT gene has low sensitivity to DNase I in fibrosarcoma cells. Insertion of the polyomavirus enhancer and neo reporter gene into exon 3 confers altered HPRT DNase I sensitivity for several kilobases on either side of the enhancer. The changes in DNase I sensitivity peak near the enhancer and decline with distance from the enhancer. The increase in HPRT DNase I sensitivity persisted when the tk promoter was deleted from the inserted construct but disappeared when the enhancer was deleted. These experiments identify the polyomavirus enhancer as a cis-acting initiator of chromatin accessibility.
Pikaart
M
M
Department of Biological Chemistry, University of Michigan, Ann Arbor 48109-2007.
Feng
J
J
Villeponteau
B
B
D001483:Base Sequence; D002460:Cell Line; D002843:Chromatin / Q000378:metabolism*; D003850:Deoxyribonuclease I; D004742:Enhancer Elements, Genetic; D006801:Humans; D007041:Hypoxanthine Phosphoribosyltransferase / Q000235:genetics* / Q000378:metabolism; D008969:Molecular Sequence Data; D016133:Polymerase Chain Reaction; D011120:Polyomavirus / Q000235:genetics*; D011401:Promoter Regions, Genetic; D015183:Restriction Mapping; D014158:Transcription, Genetic; D016662:Virus Integration / Q000235:genetics*
D002843:Chromatin; D007041:Hypoxanthine Phosphoribosyltransferase; D003850:Deoxyribonuclease I