Flow cytometric analysis of the cell cycle in polyamine-depleted cells.
Polyamines are found in all cells but their function is not fully understood. We have studied the effect of polyamines on the passage of cells through the cell cycle using a polyamine auxotrophic mutant, CHO-P22, which has no detectable ornithine decarboxylase activity. The ability of these cells to grow without serum allows efficient polyamine depletion. A flow cytometric analysis of DNA content and bromodeoxyuridine labeling showed that without added polyamines the cells accumulated in S-phase, the rate of DNA synthesis was retarded, and the entry into mitosis was blocked. Addition of polyamines to cultures deprived of polyamines induced cells in all phases of the cell cycle to reinitiate cycling. Earlier studies have shown that cells with damaged DNA are blocked from entering into mitosis but caffeine can partly overcome this block and induce premature chromosome condensation. Polyamine-depleted CHO-P22 cells responded to caffeine in the same way as cells with damaged DNA. These results show that polyamine depletion in CHO-P22 cells primarily affects DNA synthesis. The finding that polyamine-starved cells continuously take up bromodeoxyuridine without a corresponding increase in the amount of DNA is compatible with extensive repair of erroneous and/or damaged DNA. Polyamine auxotrophic Chinese hamster ovary (CHO) cells might be useful in studies on the regulation of mitosis in mammalian cells.
Pohjanpelto
P
P
Department of Virology, University of Helsinki, Finland.
Nordling
S
S
Knuutila
S
S
D000818:Animals; D001973:Bromodeoxyuridine / Q000378:metabolism; D016466:CHO Cells / Q000737:chemistry / Q000187:drug effects / Q000201:enzymology; D002110:Caffeine / Q000494:pharmacology; D002453:Cell Cycle / Q000187:drug effects; D002875:Chromosomes / Q000187:drug effects / Q000648:ultrastructure; D006224:Cricetinae; D003412:Cricetulus; D016895:Culture Media, Serum-Free; D004247:DNA / Q000032:analysis*; D004249:DNA Damage; D004260:DNA Repair; D004261:DNA Replication; D005434:Flow Cytometry; D008938:Mitosis / Q000187:drug effects; D009955:Ornithine Decarboxylase / Q000172:deficiency; D011073:Polyamines; D011700:Putrescine / Q000494:pharmacology
D016895:Culture Media, Serum-Free; D011073:Polyamines; D002110:Caffeine; D004247:DNA; D009955:Ornithine Decarboxylase; D001973:Bromodeoxyuridine; D011700:Putrescine