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Intracellular Localization of Human DNA Repair Enzyme Methylguanine-DNA Methyltransferase by Antibodies and Its Importance¹

Chemical Carcinogenesis Laboratory, Institute of Molecular and Cell Biology [T. C. A., R. B. A., B. F. L. L.], Bioprocessing Technology Unit, Department of Chemical Engineering [K. C. L.], National University of Singapore, Kent Ridge, Singapore 0511, Republic of Singapore

² To whom requests for reprints should be addressed.

The human DNA repair enzyme, methylguanine-DNA methyltransferase (MGMT, M_r 21,000), which protects cells against the mutagenic effect of alkylating carcinogens, was found to be localized in the cell nucleus (except the nucleolus) by immunofluorescence staining using polyclonal and monoclonal antibodies. The supporting experiments came from differential staining of the MGMT-deficient (mer–) and -proficient (mer+) cells, Western blotting analysis, and specific antibody depletion studies with the immobilized fusion protein, GSTMGMT-glutathione-Sepharose. Its localization in the nucleus agrees with its biological function and possibly explains the ineffective protection of mammalian cells (mer–) transfected with the Escherichia coli MGMT genes from bifunctional alkylating agents.

¹ This research is funded by the National University of Singapore.

The costs of publication of this article were defrayed in part by the payment of page charges. This article must therefore be hereby marked advertisement in accordance with 18 U.S.C. Section 1734 solely to indicate this fact.

Received 4/22/92. Accepted 9/17/92.

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