A cis-acting element accounts for a conserved methylation pattern upstream of the mouse adenine phosphoribosyltransferase gene.
A 2.1-kilobase pair region located just upstream of the mouse aprt (adenine phosphoribosyltransferase) gene has a methylation pattern that is conserved in mouse tissues and culture cell lines. This upstream region includes four HpaII/MspI sites. Two of these sites are fully methylated, one is partially methylated, and one is unmethylated. Transfection experiments have demonstrated that the conserved methylation pattern can be reproduced in a mouse embryonal carcinoma stem cell line via de novo methylation (Turker, M.S., Mummaneni, P., and Bishop, P.L. (1991) Somat. Cell Mol. Genet. 17, 151-157). To examine the molecular basis of the conserved methylation pattern, a plasmid-based deletion analysis was conducted by removing and rearranging specific portions of the upstream region. Unmethylated versions of these plasmid constructs were then transfected into the mouse stem cell line and the methylation status of the remaining HpaII/MspI sites determined with a Southern blot analysis. By using this approach, a cis-acting sequence within the upstream region of approximately 0.8 kilobase pairs was identified which appears responsible for the conserved methylation pattern. We use the term "de novo methylation center" to denote this sequence. Based on the results obtained, a model is offered to explain the formation of the conserved methylation pattern in the upstream region.
Mummaneni
P
P
Department of Pathology, University of Kentucky College of Medicine, Lexington 40536.
Bishop
P L
PL
Turker
M S
MS
D000228:Adenine Phosphoribosyltransferase / Q000235:genetics*; D000818:Animals; D015139:Blotting, Southern; D004247:DNA / Q000235:genetics* / Q000302:isolation & purification / Q000378:metabolism; D008745:Methylation; D051379:Mice; D010957:Plasmids; D012045:Regulatory Sequences, Nucleic Acid; D015183:Restriction Mapping; D017384:Sequence Deletion; D013724:Teratoma; D014162:Transfection; D014407:Tumor Cells, Cultured
D004247:DNA; D000228:Adenine Phosphoribosyltransferase