Multiple subphases of DNA repair and poly (ADP-ribose) synthesis in Chinese hamster ovary (CHO-K1) cells.
The two types of DNA synthesis as well as poly(ADP-ribose) biosynthesis were measured simultaneously in synchronized intact populations of CHO cells throughout the duration of S phase. Naturally occurring DNA fragmentation was detected by random primed oligonucleotide synthesis (ROPS assay). Fractions of synchronous cell populations were obtained by counterflow centrifugal elutriation. By gradually increasing the resolution of centrifugal elutriation multiple non-overlapping repair and replication peaks were obtained. The elutriation profile of DNA repair peaks corresponded to the DNA fragmentation pattern measured by ROPS assay. The number and position of poly(ADP-ribose) peaks during S phase resembled those seen in the DNA replication profile. Our results indicate that PAR synthesis is coupled to DNA replication serving the purpose of genomic stability.
Szepessy
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Department of Animal Anatomy and Physiology, University of Debrecen, Debrecen, Hungary.
Nagy
Gabor
G
Jenei
Zsolt
Z
Serfozo
Zoltan
Z
Csuka
Ildiko
I
James
Jill
J
Banfalvi
Gaspar
G
D000818:Animals; D016466:CHO Cells; D002452:Cell Count; D002453:Cell Cycle / Q000235:genetics; D006224:Cricetinae; D004247:DNA / Q000235:genetics / Q000378:metabolism*; D053938:DNA Fragmentation / Q000235:genetics; D004260:DNA Repair; D004261:DNA Replication / Q000235:genetics; D005434:Flow Cytometry; D011064:Poly Adenosine Diphosphate Ribose / Q000096:biosynthesis*
D011064:Poly Adenosine Diphosphate Ribose; D004247:DNA