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Blood, 1 December 2001, Vol. 98, No. 12, pp. 3441-3446
RED CELLS
De novo methylation of an embryonic globin gene during normal
development is strand specific and spreads from the proximal
transcribed region
Rakesh Singal and
Jane M. vanWert
From the Department of Medicine, Overton Brooks VA
Medical Center and Feist-Weiller Cancer Center, Louisiana State
University Health Sciences Center, Shreveport, LA.
The recently discovered de novo methyltransferases DNMT3a and
DNMT3b have been shown to be critical to embryonic development. However, at a single gene level, little is known about how the methylation pattern is established during development. The avian embryonic  -globin gene promoter is completely unmethylated in 4-day-old chicken embryonic erythroid cells, where it is expressed at a
high level, and completely methylated in adult erythroid cells, where
it is silent. The methylation pattern of the -globin gene promoter,
proximal transcribed region, and distal transcribed region on both DNA
strands was examined during development in chicken erythroid cells. It
was found that de novo methylation targets the CpG-dense proximal
transcribed region on the coding (top) strand initially, followed by
spreading into the 3' region and into the promoter region. Methylation
of the template (bottom) strand lags behind that of the coding strand,
and complete methylation of both strands occurs only after the gene has
been silenced. The results of the study indicate that establishment of
the de novo methylation pattern involves strand-specificity and
methylation spreading.
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