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Hypomethylation andADA gene expression in mouse CAK cells

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Somatic Cell and Molecular Genetics

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 ADAin 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 ADAin 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 ADAgene. Molecular analysis of the parental CAK cells and the ADA-positive derivative clones demonstrated that both 5′ and 3′ regions of the ADAgene had become hypomethylated in the ADAM + clones. We conclude that methylation is another element involved with the transcriptional control of the ADAgene and that ADAmight serve as a useful model for studying the interaction of cis-and trans-acting regulational elements.

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Author notes

  1. Raymond L. Stallings

    Present address: Genetics Group, Los Alamos National Laboratory, 87545, Los Alamos, New Mexico

Authors and Affiliations

  1. Department of Genetics, The University of Texas System Cancer Center, M.D. Anderson Hospital and Tumor Institute, Texas Medical Center, 77054, Houston, Texas

    Raymond L. Stallings & Michael J. Siciliano

  2. Department of Medical Oncology, The University of Texas System Cancer Center, M.D. Anderson Hospital and Tumor Institute, Texas Medical Center, 77030, Houston, Texas

    Marsha L. Frazier

  3. Department of Biochemistry, Baylor College of Medicine, Texas Medical Center, 77030, Houston, Texas

    Muayyad R. Al-Ubaidi & Rodney E. Kellems

Authors
  1. Raymond L. Stallings
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  2. Michael J. Siciliano
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  3. Marsha L. Frazier
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  4. Muayyad R. Al-Ubaidi
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  5. Rodney E. Kellems
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Stallings, R.L., Siciliano, M.J., Frazier, M.L. et al. Hypomethylation andADA gene expression in mouse CAK cells. Somat Cell Mol Genet 15, 1–11 (1989). https://doi.org/10.1007/BF01534664

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  • DOI: https://doi.org/10.1007/BF01534664

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