Recapitulation of the cellular xeroderma pigmentosum-variant phenotypes using short interfering RNA for DNA polymerase H.
The lesion-specific DNA polymerase POLH gene is mutated in xeroderma pigmentosum variant (XP-V) patients who exhibit an increased skin cancer incidence from UV exposure. Normal cells in which POLH expression was reduced using short interfering RNAs (siRNAs) were compared with the XP-V cellular phenotype that results from naturally occurring inactivating mutations. Stable clones expressing siRNA had partially reduced POLH protein levels, and intermediate levels of UV sensitivity and S phase checkpoint activation, but similar levels of Mre11 foci as in XP-V cells. Therefore, suppression of POLH expression levels by siRNA recapitulates most of the phenotypes seen in cells from XP-V patients with inactivating mutations in POLH.
Laposa
Rebecca R
RR
Department of Dermatology, University of California, San Francisco, California 94143-0808, USA.
Feeney
Luzviminda
L
Cleaver
James E
JE
D002110:Caffeine / Q000494:pharmacology; D002461:Cell Line, Transformed; D004259:DNA-Directed DNA Polymerase / Q000235:genetics; D004706:Endodeoxyribonucleases / Q000235:genetics; D005090:Exodeoxyribonucleases / Q000235:genetics; D005347:Fibroblasts / Q000201:enzymology / Q000502:physiology / Q000528:radiation effects; D006801:Humans; D019384:Nucleic Acid Synthesis Inhibitors; D010957:Plasmids / Q000235:genetics; D034741:RNA, Small Interfering / Q000235:genetics*; D016196:S Phase / Q000235:genetics / Q000528:radiation effects; D029701:Saccharomyces cerevisiae Proteins / Q000235:genetics; D014162:Transfection; D014466:Ultraviolet Rays / Q000009:adverse effects; D014983:Xeroderma Pigmentosum / Q000201:enzymology* / Q000235:genetics* / Q000517:prevention & control; D000099270:Y-Family DNA Polymerases
D002110:Caffeine; D004259:DNA-Directed DNA Polymerase; D004706:Endodeoxyribonucleases; D005090:Exodeoxyribonucleases; D019384:Nucleic Acid Synthesis Inhibitors; D034741:RNA, Small Interfering; D029701:Saccharomyces cerevisiae Proteins; C112338:Rad30 protein; D000099270:Y-Family DNA Polymerases; C094191:MRE11 protein, S cerevisiae