Fidelity of DNA Replication in Normal and Malignant Human Breast Cells.
Abstract
In order to continue to determine the degree to which the accumulation of mutations in breast cancer cells is due to a change in the fidelity of the cellular DNA replication machinery. We have continued and expanded our experiments utilizing the multiprotein DNA synthesome isolated from malignant, non-malignant, and normal breast cells from both tissue culture and surgically resected human breast tissue. The DNA synthesome has been extensively demonstrated to carryout full length DNA replication in vitro and has been demonstrated by our lab to accurately depict the DNA replication process as it occurs in the intact cell. By utilizing a target gene in a forward mutagenesis assay, we have observed that the DNA synthesome from malignant breast cells replicates DNA with a significantly diminished fidelity as compared to the synthesome from non-malignant or normal breast cells. Furthermore, nucleotide sequencing has demonstrated that the types of replication errors created by the DNA synthesome derived from the malignant breast cells include nucleotide insertions, deletions, and mispairs. To begin to explore the mechanisms responsible for the diminished fidelity of DNA synthesome-mediated replication, we have initiated studies to map the possible interactions of the DNA synthesome with several DNA repair proteins. Initial data suggests that at least six key DNA repair proteins are tightly associated with the DNA synthesome.
Document Details
- Document Type
- Technical Report
- Publication Date
- Aug 01, 1997
- Accession Number
- ADA335186
Entities
People
- Jennifer W Sekowski
Organizations
- University of Maryland School of Medicine