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Nucleic Acids Research, 1984, Vol. 12, No. 6 2807-2822
© 1984

MOLECULAR BIOLOGY

Methylation of satellite sequences in mouse spermatogenic and somatic DNAs

Carola Ponzetto-Zimmerman and Debra J. Wolgemuth

Department of Human Genetics and Development and The Center for Reproductive Sciences, Columbia University, College of Physicians and Surgeons 630 West 168th Street, New York, NY 10032, USA

Received December 8, 1983. Revised February 13, 1984. Accepted February 13, 1984.

The distribution of 5-methyl cytosine (5-MeC) residues in a highly repetitive sequence, mouse major satellite, was examined in germinal versus somatic DNAs by digestion with the methylation sensitive isoschizomers Msp I and Hpa II and Southern blot analysis, using a cloned satellite probe. DNA from liver, brain, and a mouse fibroblast cell line, C3H 10T1/2, yielded a multimeric hybridization pattern after digestion with Msp I (and control Eco RI) but were resistant to digestion with Hpa II, reflecting a high level of methylation of the satellite sequences. In contrast, DNA from mature sperm was undermethylated at these same sequences as indicated by the ability of Hpa II to generate a multimeric pattern. DNAs from purified populations of testis cells in different stages of spermatogenesis were examined to determine when during germ cell differentiation the undermethylation was established. As early as in primitive type A, type A, and type B spermatogonia, an undermethylation of satellite sequences was observed. This suggests that this highly specific undermethylation of germ cell satellite DNA occurs very early in the germ cell lineage, prior to entry into meiosis


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