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Nucleic Acids Research, 2000, Vol. 28, No. 10 2108-2113
© 2000 Oxford University Press

Differential mRNA expression of the human DNA methyltransferases (DNMTs) 1, 3a and 3b during the G0/G1 to S phase transition in normal and tumor cells

Keith D. Robertson, Khandan Keyomarsi1, Felicidad A. Gonzales, Mihaela Velicescu and Peter A. Jones*

Norris Comprehensive Cancer Center, University of Southern California, MS 83, 1441 Eastlake Avenue, Los Angeles, CA 90033, USA and 1Wadsworth Center, NYS Department of Health, Room C-400, Albany, NY 12201, USA

DNA methylation is essential for mammalian development, X-chromosome inactivation, and imprinting yet aberrant methylation patterns are one of the most common features of transformed cells. One of the proposed causes for these defects in the methylation machinery is overexpression of one or more of the three known catalytically active DNA methyltransferases (DNMTs) 1, 3a and 3b, yet there are clearly examples in which overexpression is minimal or non-existent but global methylation anomalies persist. An alternative mechanism which could give rise to global methylation errors is the improper expression of one or more of the DNMTs during the cell cycle. To begin to study the latter possibility we examined the expression of the mRNAs for DNMT1, 3a and 3b during the cell cycle of normal and transformed cells. We found that DNMT1 and 3b levels were significantly downregulated in G0/G1 while DNMT3a mRNA levels were less sensitive to cell cycle alterations and were maintained at a slightly higher level in tumor lines compared to normal cell strains. Enzymatic activity assays revealed a similar decrease in the overall methylation capacity of the cells during G0/G1 arrest and again revealed that a tumor cell line maintained a higher methylation capacity during arrest than a normal cell strain. These results reveal a new level of control exerted over the cellular DNA methylation machinery, the loss of which provides an alternative mechanism for the genesis of the aberrant methylation patterns observed in tumor cells.

* To whom correspondence should be addressed. Tel: +1 323 865 0816; Fax +1 323 865 0102; Email: jones_p@ccnt.hsc.usc.edu Present address: Keith D. Robertson, LME/NICHD/NIH, Building 18T, Room 106, 18 Library Drive, Bethesda, MD 20892, USA


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