Large-scale chromatin decondensation and recondensation regulated by transcription from a natural promoter

doi:10.1083/jcb.200011069

Published 9 July 2001. doi:10.1083/jcb.200011069

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© The Rockefeller University Press, 0021-9525/2001/7/33 $5.00
The Journal of Cell Biology, Volume 154, Number 1, July 9, 2001 33-48

Article

Large-scale chromatin decondensation and recondensation regulated by transcription from a natural promoter

Waltraud G. Müller, Dawn Walker, Gordon L. Hager and James G. McNally

Laboratory of Receptor Biology and Gene Expression, National Cancer Institute, National Institutes of Health, Bethesda, MD 20892

Address correspondence to James G. McNally, Laboratory of Receptor Biology and Gene Expression, National Institutes of Health, Bldg. 41, Rm. B-602, 41 Library Dr., MSC 5055, Bethesda, MD 20892-5055. Tel.: (301) 402-0209. Fax: (301) 496-4951. E-mail: mcnallyj{at}exchange.nih.gov

We have examined the relationship between transcription andchromatin structure using a tandem array of the mouse mammarytumor virus (MMTV) promoter driving a ras reporter. The arraywas visualized as a distinctive fluorescent structure in livecells stably transformed with a green fluorescent protein (GFP)-taggedglucocorticoid receptor (GR), which localizes to the repeatedMMTV elements after steroid hormone treatment. Also found atthe array by immunofluorescence were two different steroid receptorcoactivators (SRC1 and CBP) with acetyltransferase activity,a chromatin remodeler (BRG1), and two transcription factors(NFI and AP-2). Within 3 h after hormone addition, arrays visualizedby GFP-GR or DNA fluorescent in situ hybridization (FISH) decondensedto varying degrees, in the most pronounced cases from a $~$ 0.5-µmspot to form a fiber 1–10 µm long. Arrays laterrecondensed by 3–8 h of hormone treatment. The degreeof decondensation was proportional to the amount of transcriptproduced by the array as detected by RNA FISH. Decondensationwas blocked by two different drugs that inhibit polymerase II,5,6-dichloro-1-ß-D-ribofuranosylbenzimidazole (DRB)and ${alpha}$ -amanitin. These observations demonstrate a role for polymerasein producing and maintaining decondensed chromatin. They alsosupport fiber-packing models of higher order structure and suggestthat transcription from a natural promoter may occur at muchhigher DNA-packing densities than reported previously.

Key Words: higher order chromatin structure; transcription; glucocorticoid receptor

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