Altered DNA polymerase iota expression in breast cancer cells leads to a reduction in DNA replication fidelity and a higher rate of mutagenesis.
The recently discovered human enzyme DNA polymerase iota (pol iota) has been shown to have an exceptionally high error rate on artificial DNA templates. Although there is a considerable body of in vitro evidence for a role for pol iota in DNA lesion bypass, there is no in vivo evidence to confirm this action. We report here that pol iota expression is elevated in breast cancer cells and correlates with a significant decrease in DNA replication fidelity. We also demonstrate that UV treatment of breast cancer cells additionally increases pol iota expression with a peak occurring between 30 min and 2 h after cellular insult. This implies that the change in pol iota expression is an early event after UV-mediated DNA damage. That pol iota may play a role in the higher mutation frequencies observed in breast cancer cells was suggested when a reduction in mutation frequency was found after pol iota was immunodepleted from nuclear extracts of the cells. Analysis of the UV-induced mutation spectra revealed that > 90% were point mutations. The analysis also demonstrated a decreased C --> T nucleotide transition and an increased C --> A transversion rate. Overall, our data strongly suggest that pol iota may be involved in the generation of both increased spontaneous and translesion mutations during DNA replication in breast cancer cells, thereby contributing to the accumulation of genetic damage.
Department of Medicine, Division of Hematology/Oncology, Cancer Research Institute, Indiana University of School of Medicine, Indianapolis, Indiana 46202, USA.
Jin
J
Yang
Zhiwen
Z
Chen
Yang
Y
Liu
Robert J
RJ
Hickey
Linda H
LH
Malkas
D001483:Base Sequence; D001940:Breast / Q000166:cytology / Q000201:enzymology / Q000502:physiology / Q000528:radiation effects; D001943:Breast Neoplasms / Q000201:enzymology* / Q000235:genetics*; D045744:Cell Line, Tumor; D004249:DNA Damage; D004260:DNA Repair; D004261:DNA Replication / Q000502:physiology*; D004273:DNA, Neoplasm / Q000528:radiation effects; D004259:DNA-Directed DNA Polymerase / Q000096:biosynthesis* / Q000528:radiation effects; D004790:Enzyme Induction / Q000528:radiation effects; D006801:Humans; D008969:Molecular Sequence Data; D016296:Mutagenesis / Q000502:physiology* / Q000528:radiation effects; D017354:Point Mutation; D000094769:DNA Polymerase iota
D004273:DNA, Neoplasm; D004259:DNA-Directed DNA Polymerase; D000094769:DNA Polymerase iota; C000722070:POLI protein, human