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Nucleic Acids Research, 1982, Vol. 10, No. 11 3459-3474
© 1982

MOLECULAR BIOLOGY

Relative levels of methylation in human growth hormone and chorionic somatomammotropin genes in expressing and non-expressing tissues

Brian L. Hjelle*, John A. Phillips, III* and Peter H. Seeburg{dagger}

*Department of Pediatrics, John Hopkins University School of Medicine Baltimore, MD 21205 {dagger}Department of Molecular Biology Genentech Inc., South San Francisco, CA 94080, USA

Received December 28, 1981. Revised March 29, 1982. Accepted May 13, 1982.

It has been shown that the extent of methylation of cytosine in vertebrate DNA is inversely correlated with gene expression. We studied cytosine methylation in and around the homologous human growth hormone (GH) and chorionic somatomammotropin (CS) genes to determine if these genes are undermethylated in DNA from tissues in which they are expressed (pituitary and placenta, respectively) compared to other tissues. Hpa II and Hha I (which cleave only unmethylated 5' CCGG 3' and 5' GCGC 3' respectively) and Msp I (which cleaves CCGG and Cme CGG) were used to digest DNA samples followed by gel electrophoresis, Southern transfer and hybridization with a GH cDNA probe. The extent of methylation of Hpa II and Hha I sites in the GH and CS genes was leukocyte>>pituitary>placenta=hydatidiform mole. Taken as a whole, our data support the hypothesis that undermethylation is a necessary but not sufficient condition for gene expression since placental and pituitary DNAs are less rnethylated than leukocyte DNA in this region. However, the correlation between gene expression and undermethylation is imperfect since (1) hydatiform mole DNA has a very similar methylation pattern compared to placental DNA even though moles make little or no CS and (2) the level of methylation of the GH gene compared to the CS gene does not vary in a tissue-specific manner.


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