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Molecular and Cellular Biology, January 1999, p. 164-172, Vol. 19, No. 1
0270-7306/99/$04.00+0
Copyright © 1999, American Society for Microbiology. All rights reserved.

DNA Methylation Profile of the Mouse Skeletal alpha -Actin Promoter during Development and Differentiation

Peter M. Warnecke1,2 and Susan J. Clark1,3,*

Kanematsu Laboratories, Royal Prince Alfred Hospital, Camperdown, New South Wales 2050,1 School of Biological Sciences, University of Sydney, New South Wales 2006,2 and CSIRO Division of Molecular Science, Sydney Laboratory, North Ryde, New South Wales 2113,3 Australia

Received 4 August 1998/Returned for modification 9 September 1998/Accepted 17 September 1998

Genomic levels of DNA methylation undergo widespread alterations in early embryonic development. However, changes in embryonic methylation have proven difficult to study at the level of single-copy genes due to the small amount of tissue available for assay. This study provides the first detailed analysis of the methylation state of a tissue-specific gene through early development and differentiation. Using bisulfite sequencing, we mapped the methylation profile of the tissue-specific mouse skeletal alpha -actin promoter at all stages of development, from gametes to postimplantation embryos. We show that the alpha -actin promoter, which is fully methylated in the sperm and essentially unmethylated in the oocyte, undergoes a general demethylation from morula to blastocyst stages, although the blastula is not completely demethylated. Remethylation of the alpha -actin promoter occurs after implantation in a stochastic pattern, with some molecules being extensively methylated and others sparsely methylated. Moreover, we demonstrate that tissue-specific expression of the skeletal alpha -actin gene in the adult mouse does not correlate with the methylation state of the promoter, as we find a similar low level of methylation in both expressing and one of the two nonexpressing tissues tested. However, a subset of CpG sites within the skeletal alpha -actin promoter are preferentially methylated in liver, a nonexpressing tissue.

* Corresponding author. Mailing address: CSIRO Division of Molecular Science, Sydney Laboratory, 2 Richardson Place, Riverside Corporate Park, Delhi Rd., North Ryde, NSW 2113, Australia. Phone: (612) 94905148. Fax: (612) 94905005. E-mail: susan.clark{at}molsci.csiro.au.

Molecular and Cellular Biology, January 1999, p. 164-172, Vol. 19, No. 1
0270-7306/99/$04.00+0
Copyright © 1999, American Society for Microbiology. All rights reserved.


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