Isolation of Chinese hamster ovary cells with reduced poly(ADP-ribose) polymerase activity.
The biological function of poly(ADP-ribose) polymerase in DNA repair, cell-cycle regulation and cellular differentiation has yet to be defined. Isolation of cells which are deficient in poly(ADP-ribose) synthesis would greatly facilitate the determination of the biological role of this enzyme. A method is described for isolating Chinese hamster ovary (CHO) cells deficient in the poly(ADP-ribose) polymerase activity by direct screening of colonies for enzyme activity. Colonies with decreased production of poly(ADP-ribose) are recovered from nylon replicas for further analysis. Using this method we have isolated a series of CHO cells which have 50% or less poly(ADP-ribose) polymerase activity. These mutants have normal generation times and are 20% more sensitive to the effects of DNA (m)ethylating agents than the parental cell. However, these mutants display normal sensitivity to gamma-rays.
MacLaren
R A
RA
Wistar Institute of Anatomy and Biology, Philadelphia, PA 19104.
Witmer
M V
MV
Richardson
E
E
Stamato
T D
TD
D000818:Animals; D002455:Cell Division; D002460:Cell Line; D002470:Cell Survival; D006224:Cricetinae; D005020:Ethyl Methanesulfonate / Q000633:toxicity; D007700:Kinetics; D009154:Mutation; D011065:Poly(ADP-ribose) Polymerases / Q000235:genetics / Q000378:metabolism*
D005020:Ethyl Methanesulfonate; D011065:Poly(ADP-ribose) Polymerases