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Biallelic Methylation and Silencing of Mouse Aprt in Normal Kidney Cells¹

Center for Research on Occupational and Environmental Toxicology, Oregon Health Sciences University, Portland, Oregon 97201 [J. A. R., P. A. Y., M. S. T.]; Department of Pathology and Laboratory Medicine, University of Kentucky, Lexington, Kentucky 40536 [J. S.]; Department of Genetics, Rutgers, the State University of New Jersey, Piscataway, New Jersey 08854 [J. A. T.]; and Department of Cell Biology, Neurobiology, and Anatomy, University of Cincinnati, College of Medicine, Cincinnati, Ohio 45267-0521 [P. J. S.]

Heritable gene silencing is an important mechanism of tumor suppressor gene inactivation in a variety of human cancers. In the present study, we show that methylation-associated silencing of the autosomal adenine phosphoribosyltransferase (Aprt) locus occurs in primary mouse kidney cells. Aprt-deficient cells were isolated from mice that were heterozygous for Aprt, i.e., they contained one wild-type Aprt allele and one targeted allele bearing an insertion of the bacterial neo gene. Although silencing of the wild-type allele alone was sufficient for the cells to become completely Aprt-deficient, biallelic methylation of the promoter region was found to occur. Moreover, despite the absence of selective pressure against the targeted allele, phenotypic silencing of the inserted neo gene accompanied silencing of the wild-type Aprt allele. A potential role for allelic homology in these events is discussed.

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