Nucleic Acids Research, 2000, Vol. 28, No. 13 2541-2550
© 2000 Oxford University Press
HMG I/Y regulates long-range enhancer-dependent transcription on DNA and chromatin by changes in DNA topology
Regulatory Biology Laboratory, The Salk Institute for Biological Studies, 10010 North Torrey Pines Road, La Jolla, CA 92037, USA and 1Lineberger Comprehensive Cancer Center, University of North Carolina, Chapel Hill, NC 27599, USA
The nature of nuclear structures that are required to confer transcriptional regulation by distal enhancers is unknown. We show that long-range enhancer-dependent ß-globin transcription is achieved in vitro upon addition of the DNA architectural protein HMG I/Y to affinity-enriched holo RNA polymerase II complexes. In this system, HMG I/Y represses promoter activity in the absence of an associated enhancer and strongly activates transcription in the presence of a distal enhancer. Importantly, nucleosome formation is neither necessary for long-range enhancer regulation in vitro nor sufficient without the addition of HMG I/Y. Thus, the modulation of DNA structure by HMG I/Y is a critical regulator of long-range enhancer function on both DNA and chromatin-assembled genes. Electron microscopic analysis reveals that HMG I/Y binds cooperatively to preferred DNA sites to generate distinct looped structures in the presence or absence of the ß-globin enhancer. The formation of DNA topologies that enable distal enhancers to strongly regulate gene expression is an intrinsic property of HMG I/Y and naked DNA.
* To whom correspondence should be addressed. Tel: +1 858 453 6560; Fax: +1 858 535 8194; Email: emerson@salk.edu
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