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Vol. 13, No. 22, pp. 3003-3014, November 15, 1999
1 Howard Hughes Medical Institute and Departments of
Microbiology and Biochemistry, University of San Francisco,
San Francisco, California 94143 USA
The immunoglobulin intragenic µ enhancer region acts as a locus
control region that mediates transcriptional activation over large
distances in germ line transformation assays. In transgenic mice, but
not in transfected tissue culture cells, the activation of a variable
region (VH) promoter by the µ enhancer is dependent on flanking nuclear matrix attachment regions (MARs). Here, we examine
the effects of DNA methylation, which occurs in early mouse
development, on the function of the µ enhancer and the MARs. We find
that methylation of rearranged µ genes in vitro, before transfection,
represses the ability of the µ enhancer to activate the
VH promoter over the distance of 1.2 kb. However,
methylation does not affect enhancer-mediated promoter activation over
a distance of 150 bp. In methylated DNA templates, the µ enhancer
alone induces only local chromatin remodeling, whereas in combination
with MARs, the µ enhancer generates an extended domain of histone
acetylation. These observations provide evidence that DNA methylation
impairs the distance independence of enhancer function and thereby
imposes a requirement for additional regulatory elements, such as MARs, which facilitate long-range chromatin remodeling.
[Key Words: MAR; enhancer; LCR; methylation; chromatin]
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