Abstract
Mercuric ion is cytotoxic and mutagenic to cells; however, the mechanisms of mercuric ion-induced cytotoxicity are not well understood. Numerous studies have suggested that these effects may be due in part to the alteration and inhibition of a variety of cellular processes including DNA replication, DNA repair, RNA transcription, and protein synthesis. Studies utilizing whole cells to examine these activities are not able to specifically identify the precise mechanism or site of the effect. Other studies carried out using whole cell extracts and variously purified DNA polymerases are not able to adequately represent the highly ordered environment in which DNA replication occurs in the intact cell. We report here, for the first time, the use of an intact human cell multiprotein complex (which we have termed the DNA synthesome) to carry out full-length DNA replication and DNA synthesis in the presence of Hg2+ionin vitro.In this study we report that DNA replication and DNA polymerase activity, as well as DNA replication fidelity of the human cell DNA synthesome, are specifically inhibited by physiologically attainable concentrations of mercuric ion.
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