title: Hypomethylation and ADA gene expression in mouse CAK cells.
abstract: The adenosine deaminase (ADA) locus appears to be under complex transcriptional control since levels of ADA enzyme activity vary greatly between different tissues and stages of development. Evidence that a trans-acting factor may be involved with the regulation of this locus came from previous experiments where fusion of ADA-negative human JEG cells and mouse ADA-positive cells led to the trans-activation of human ADA in a hybrid nucleus. Here, we demonstrate that the near euploid mouse embryo fibroblast cell line, CAK, also lacks detectable ADA enzyme activity due to altered gene regulation. We further demonstrate that ADA in CAK cells is not amenable to activation by somatic cell fusion. Following treatment with 5-azacytidine and Xyl-A selection (for ADA), however, CAK clones were obtained that stably express the ADA gene. Molecular analysis of the parental CAK cells and the ADA-positive derivative clones demonstrated that both 5' and 3' regions of the ADA gene had become hypomethylated in the ADA+ clones. We conclude that methylation is another element involved with the transcriptional control of the ADA gene and that ADA might serve as a useful model for studying the interaction of cis- and trans-acting regulational elements.
authors: Department of Genetics, University of Texas System Cancer Center, M.D. Anderson Hospital and Tumor Institute, Texas Medical Center, Houston 77054.
authors: R L
authors: RL
authors: Stallings
authors: M J
authors: MJ
authors: Siciliano
authors: M L
authors: ML
authors: Frazier
authors: M R
authors: MR
authors: al-Ubaidi
authors: R E
authors: RE
authors: Kellems
mesh_terms: D000243:Adenosine Deaminase / Q000096:biosynthesis / Q000235:genetics*; D000818:Animals; D001374:Azacitidine / Q000494:pharmacology; D015152:Blotting, Northern; D015139:Blotting, Southern; D002460:Cell Line; D006224:Cricetinae; D004247:DNA / Q000378:metabolism*; D005786:Gene Expression Regulation; D006822:Hybrid Cells; D007527:Isoenzymes / Q000096:biosynthesis; D008745:Methylation; D051379:Mice; D009700:Nucleoside Deaminases / Q000235:genetics*; D014157:Transcription Factors / Q000235:genetics*
chemical_list: D007527:Isoenzymes; D014157:Transcription Factors; D004247:DNA; D009700:Nucleoside Deaminases; D000243:Adenosine Deaminase; D001374:Azacitidine