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Nucleic Acids Research, 1980, Vol. 8, No. 20 4563-4574
© 1980

MOLECULAR BIOLOGY

A novel type of secondary modification of two CCGG residues in the human {gamma}{delta}ß-globin gene locus

L. H. T.van der Ploeg*, J. Groffen+ and R.A. Flavell+,*

*Section for Medical Enzymology and Molecular Biology, Lab. Biochemistry, University of Amsterdam, Jan Swammerdam Inst. 1005 GA Amsterdam, Netherlands +Lab. Gene Structure and Expression, National Inst. Medical Research Mill Hill, London NW7 1AA, UK

Received September 8, 1980. The majority of the CCGG residues in the human {gamma}{delta}ß-globin gene locus are cleaved by Map I, irrespective of the tissue of origin of the DNA, although these sites show differential sensitivity to Hap II as a result of methylation of the internal C residue of the cleavage site (ref. 6). Two CCGC sites, at homologous positions 54 nucleotides in front of the G{gamma}- and A{gamma}-globin genes respectively, are not cleaved by Msp I in DNA from several human tissues, although DNA from placenta, foetal liver and from some established cell lines is cut at these sites. We have cloned the A{gamma}-globin gene from foetal blood DNA where the relevant CCGG site is not cut by Map I. After cloning, the CCGG Site can be cut by Msp I. The failure to cleave at this CCGG site in foetal blood DNA therefore, is not the result of a change in the DNA sequence of the cleavage site. Most likely the external C residue and perhaps both C residues are blocked by methylation at these two specific sites.


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