'Knock down' of DNA polymerase beta by RNA interference: recapitulation of null phenotype.
DNA polymerase beta (pol beta) is the major DNA polymerase involved in the base excision repair (BER) pathway in mammalian cells and, as a consequence, BER is severely compromised in cells lacking pol beta. Pol beta null (-/-) mouse embryos are not viable and pol beta null cells are hypersensitive to alkylating agents. Using RNA interference (RNAi) technology in mouse cells, we have reduced the pol beta protein and mRNA to undetectable levels. Pol beta knockdown cell lines display a pattern of hypersensitivity to DNA damaging agents similar to that observed in pol beta null cells. Generation of pol beta knock down cells makes it possible to combine the pol beta null phenotype with deficiencies in other DNA repair proteins, thereby helping to elucidate the role of pol beta and its interactions with other proteins in mammalian cells.
Polosina
Yaroslava Y
YY
Laboratory of Chemical Biology, Department of Pharmacological Sciences, State University of New York, Stony Brook, NY 11794, USA.
Rosenquist
Thomas A
TA
Grollman
Arthur P
AP
Miller
Holly
H
D000818:Animals; D001483:Base Sequence; D001761:Bleomycin / Q000633:toxicity; D002460:Cell Line; D002945:Cisplatin / Q000633:toxicity; D004249:DNA Damage; D019951:DNA Polymerase beta / Q000172:deficiency* / Q000235:genetics* / Q000378:metabolism; D004260:DNA Repair; D018076:DNA, Complementary / Q000235:genetics; D006861:Hydrogen Peroxide / Q000633:toxicity; D008741:Methyl Methanesulfonate / Q000633:toxicity; D008751:Methylene Blue / Q000633:toxicity; D051379:Mice; D010641:Phenotype; D034622:RNA Interference; D012333:RNA, Messenger / Q000235:genetics / Q000378:metabolism; D014466:Ultraviolet Rays / Q000009:adverse effects
D018076:DNA, Complementary; D012333:RNA, Messenger; D001761:Bleomycin; D008741:Methyl Methanesulfonate; D006861:Hydrogen Peroxide; D019951:DNA Polymerase beta; D002945:Cisplatin; D008751:Methylene Blue