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Molecular and Cellular Biology, May 2001, p. 3416-3424, Vol. 21, No. 10
0270-7306/01/$04.00+0   DOI: 10.1128/MCB.21.10.3416-3424.2001
Copyright © 2001, American Society for Microbiology. All rights reserved.

Protein Binding Protects Sites on Stable Episomes and in the Chromosome from De Novo Methylation

Li Han, Iping G. Lin, and Chih-Lin Hsieh*

Department of Urology and Department of Biochemistry and Molecular Biology, University of Southern California, Los Angeles, California 90033

Received 16 January 2001/Accepted 19 February 2001

We have utilized the Escherichia coli lac repressor-operator system to test whether protein binding can interfere with de novo DNA methylation in mammalian cells. We find that a DNA binding protein can protect sites on the episome as well as in the genome from the de novo methylation activity of Dnmt3a. Transcriptional machinery moving through the binding sites does not affect the de novo methylation of these sites, and it does not affect the binding protein protection of these sites from de novo methylation. This study and previous studies provide a possible mechanism for the observation that an Sp1 site can serve as a cis-acting signal for demethylation and for preventing de novo methylation of the CpG island upstream of the mouse adenine phosphoribosyltransferase (Aprt) gene. These findings also support the hypothesis that protein binding may play a crucial role in changes of CpG methylation pattern in mammalian cells.

* Corresponding author. Mailing address: Department of Urology and Department of Biochemistry and Molecular Biology, University of Southern California, 1441 Eastlake Ave., Rm. 5420, Norris Cancer Center, Mail Stop 73, Los Angeles, CA 90033. Phone: (323) 865-0567. Fax: (323) 865-3019. E-mail: hsieh_c{at}ccnt.hsc.usc.edu.

Molecular and Cellular Biology, May 2001, p. 3416-3424, Vol. 21, No. 10
0270-7306/01/$04.00+0   DOI: 10.1128/MCB.21.10.3416-3424.2001
Copyright © 2001, American Society for Microbiology. All rights reserved.


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