Kwoh, T. J., Kwoh, D. Y., McCue, A. W., Davis, G. R., Patrick, D., Gingeras, T. R. (October 1986) Introduction and expression of the bacterial PaeR7 methylase gene in mammalian cells. Proc Natl Acad Sci U S A, 83 (20). pp. 7713-7. ISSN 0027-8424 (Print)0027-8424 (Linking)
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Abstract
An approach is devised for studying the role of DNA methylation in eukaryotic gene expression. The approach is based on the expression of site-specific bacterial methylase genes in animal cells. A model system using the cloned PaeR7 (an isoschizomer of Xho I) methylase gene was constructed to test the feasibility of this approach. Expression plasmids for the PaeR7 methylase gene were introduced into mouse Ltk- cells by cotransfection with the cloned chicken thymidine kinase (tk) gene. Several of the cell strains derived from Tk+ colonies were found to express the PaeR7 gene as judged by four criteria: the cellular DNA of these strains showed increased resistance to cleavage by Xho I; these strains contained cellular proteins that comigrated with pure PaeR7 methylase protein, as visualized by immunoblotting; PaeR7 methylase activity was found in vitro in crude extracts of total cellular protein from these strains; and murine adenovirus genomes grown on cells expressing PaeR7 methylase showed resistance to cleavage to PaeR7 endonuclease. The potential applications of this approach for the study of cellular and viral gene regulation, DNA repair, and restriction modification are discussed.
| Item Type: | Paper |
|---|---|
| Uncontrolled Keywords: | Animals Cell Line DNA metabolism DNA Cytosine 5 Methyltransferase analysis genetics DNA Repair DNA Viral metabolism Genes Bacterial Methylation Mice Nucleic Acid Hybridization Plasmids Transfection |
| Subjects: | bioinformatics > genomics and proteomics > genetics & nucleic acid processing > DNA, RNA structure, function, modification > DNA methylation bioinformatics > genomics and proteomics > genetics & nucleic acid processing > DNA, RNA structure, function, modification > genes, structure and function > gene expression |
| CSHL Authors: | |
| Communities: | CSHL labs > Gingeras lab |
| Depositing User: | CSHL Librarian |
| Date: | October 1986 |
| Date Deposited: | 14 Mar 2012 14:02 |
| Last Modified: | 09 Nov 2017 20:50 |
| PMCID: | PMC386791 |
| Related URLs: | |
| URI: | https://repository.cshl.edu/id/eprint/25237 |
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