A role for polymerase eta in the cellular tolerance to cisplatin-induced damage.
Mutation of the POLH gene encoding DNA polymerase eta (pol eta) causes the UV-sensitivity syndrome xeroderma pigmentosum-variant (XP-V) which is linked to the ability of pol eta to accurately bypass UV-induced cyclobutane pyrimidine dimers during a process termed translesion synthesis. Pol eta can also bypass other DNA damage adducts in vitro, including cisplatin-induced intrastrand adducts, although the physiological relevance of this is unknown. Here, we show that independent XP-V cell lines are dramatically more sensitive to cisplatin than the same cells complemented with functional pol eta. Similar results were obtained with the chemotherapeutic agents, carboplatin and oxaliplatin, thus revealing a general requirement for pol eta expression in providing tolerance to these platinum-based drugs. The level of sensitization observed was comparable to that of XP-A cells deficient in nucleotide excision repair, a recognized and important mechanism for repair of cisplatin adducts. However, unlike in XP-A cells, the absence of pol eta expression resulted in a reduced ability to overcome cisplatin-induced S phase arrest, suggesting that pol eta is involved in translesion synthesis past these replication-blocking adducts. Subcellular localization studies also highlighted an accumulation of nuclei with pol eta foci that correlated with the formation of monoubiquitinated proliferating cell nuclear antigen following treatment with cisplatin, reminiscent of the response to UV irradiation and further indicating a role for pol eta in dealing with cisplatin-induced damage. Together, these data show that pol eta represents an important determinant of cellular responses to cisplatin, which could have implications for acquired or intrinsic resistance to this key chemotherapeutic agent.
Albertella
Mark R
MR
KuDOS Pharmaceuticals Ltd., Cambridge Science Park, Cambridge, UK.
Green
Catherine M
CM
Lehmann
Alan R
AR
O'Connor
Mark J
MJ
D000970:Antineoplastic Agents / Q000494:pharmacology; D002453:Cell Cycle / Q000187:drug effects; D002460:Cell Line; D002470:Cell Survival / Q000187:drug effects / Q000502:physiology; D002945:Cisplatin / Q000494:pharmacology*; D004247:DNA / Q000187:drug effects / Q000378:metabolism; D004249:DNA Damage / Q000502:physiology*; D019951:DNA Polymerase beta / Q000378:metabolism; D004260:DNA Repair / Q000502:physiology; D004259:DNA-Directed DNA Polymerase / Q000172:deficiency / Q000378:metabolism / Q000502:physiology*; D019008:Drug Resistance, Neoplasm; D005347:Fibroblasts / Q000187:drug effects; D006801:Humans; D018809:Proliferating Cell Nuclear Antigen / Q000378:metabolism; D025801:Ubiquitin / Q000378:metabolism; D014983:Xeroderma Pigmentosum / Q000473:pathology; D051760:Xeroderma Pigmentosum Group A Protein / Q000502:physiology; D000099270:Y-Family DNA Polymerases
D000970:Antineoplastic Agents; D002945:Cisplatin; D004247:DNA; D019951:DNA Polymerase beta; D004259:DNA-Directed DNA Polymerase; D018809:Proliferating Cell Nuclear Antigen; D025801:Ubiquitin; D051760:Xeroderma Pigmentosum Group A Protein; C497314:XPA protein, human; C112338:Rad30 protein; D000099270:Y-Family DNA Polymerases