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J Biol Chem, Vol. 274, Issue 51, 36357-36361, December 17, 1999
From the A cis-acting methylation center that signals
de novo DNA methylation is located upstream of the mouse
Aprt gene. In the current study, two approaches were taken
to determine if tandem B1 repetitive elements found at the 3' end of
the methylation center contribute to the methylation signal. First,
bisulfite genomic sequencing demonstrated that CpG sites within the B1
elements were methylated at relative levels of 43% in embryonal stem
cells deficient for the maintenance DNA methyltransferase when compared
with wild type embryonal stem cells. Second, the ability of the B1
elements to signal de novo methylation upon stable
transfection into mouse embryonal carcinoma cells was examined. This
approach demonstrated that the B1 elements were methylated de
novo to a high level in the embryonal carcinoma cells and that
the B1 elements acted synergistically. The results from these
experiments provide strong evidence that the tandem B1 repetitive
elements provide a significant fraction of the methylation center
signal. By extension, they also support the hypothesis that one role
for DNA methylation in mammals is to protect the genome from expression
and transposition of parasitic elements.
Tandem B1 Elements Located in a Mouse Methylation Center Provide
a Target for de Novo DNA Methylation*
,, and§
Center for Research on Occupational and
Environmental Toxicology and § Department of Molecular and
Medical Genetics, Oregon Health Sciences University, Portland, Oregon
97201 and ¶ Section on Neuroendocrinology, Laboratory of
Developmental Neurobiology, 49/5A38, NICHD, National Institutes of
Health, Bethesda, Maryland 20892
*
This work was supported by a grant from the Council for
Tobacco Research (to M. S. T.) and National Institutes of Health
predoctoral training Grant T32 (to R. W. B.).The costs of publication of this article were defrayed in part by the
payment of page charges. The article
must therefore be hereby marked
"advertisement" in accordance with 18 U.S.C. Section
1734 solely to indicate this fact.
To whom correspondence should be addressed: CROET, L606,
Oregon Health Sciences University, 3181 SW Sam Jackson Park Rd., Portland, OR 97201. Tel.: 503-494-2168; Fax: 503-494-3849; E-mail turkerm@ohsu.edu.
Copyright © 1999 by The American Society for Biochemistry and Molecular Biology, Inc.
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