Large-scale chromatin structure of inducible genes: transcription on a condensed, linear template.
The structure of interphase chromosomes, and in particular the changes in large-scale chromatin structure accompanying transcriptional activation, remain poorly characterized. Here we use light microscopy and in vivo immunogold labeling to directly visualize the interphase chromosome conformation of 1-2 Mbp chromatin domains formed by multi-copy BAC transgenes containing 130-220 kb of genomic DNA surrounding the DHFR, Hsp70, or MT gene loci. We demonstrate near-endogenous transcription levels in the context of large-scale chromatin fibers compacted nonuniformly well above the 30-nm chromatin fiber. An approximately 1.5-3-fold extension of these large-scale chromatin fibers accompanies transcriptional induction and active genes remain mobile. Heat shock-induced Hsp70 transgenes associate with the exterior of nuclear speckles, with Hsp70 transcripts accumulating within the speckle. Live-cell imaging reveals distinct dynamic events, with Hsp70 transgenes associating with adjacent speckles, nucleating new speckles, or moving to preexisting speckles. Our results call for reexamination of classical models of interphase chromosome organization.
Hu
Yan
Y
Department of Cell and Developmental Biology, University of Illinois, Urbana, IL 61801, USA.
Kireev
Igor
I
Plutz
Matt
M
Ashourian
Nazanin
N
Belmont
Andrew S
AS
D000818:Animals; D002843:Chromatin / Q000648:ultrastructure*; D022202:Chromosomes, Artificial, Bacterial / Q000648:ultrastructure; D018840:HSP70 Heat-Shock Proteins / Q000235:genetics; D006801:Humans; D007150:Immunohistochemistry; D017404:In Situ Hybridization, Fluorescence; D008668:Metallothionein / Q000235:genetics; D051379:Mice; D013762:Tetrahydrofolate Dehydrogenase / Q000235:genetics; D014158:Transcription, Genetic / Q000502:physiology*; D015533:Transcriptional Activation; D019076:Transgenes
D002843:Chromatin; D018840:HSP70 Heat-Shock Proteins; D008668:Metallothionein; D013762:Tetrahydrofolate Dehydrogenase