The relationship between the rate of entry into S phase, concentration of DNA polymerase alpha, and cell volume in human diploid fibroblast-like monokaryon cells.
We have examined the kinetic relationship between the rate of entry into the S phase in human diploid fibroblast-like (HDFL) monokaryon cells and (1) the concentration of DNA polymerase alpha activity and (2) the cell volume. In the former studies, a first-order dependence between the rate of entry into the S phase and the concentration of DNA polymerase alpha activity was observed, consistent with the enzyme, or a coregulated factor, being rate limiting for this metabolic process. Examination of the nature of the dependence of the rate of entry into the S phase upon cell volume revealed a more complex relationship. The results obtained in studies with synchronized cultures are consistent with the presence of two to three rate-limiting reactants when cell volume is the independent variable. Studies with asynchronous HDFL cell cultures revealed that the smallest cells in the G1 population, presumably the early G1 cells, enter the S phase at an increasing rate as a function of cell volume up to a certain size, beyond which the cells enter at a decreasing rate similar to that observed in the studies with the synchronized cultures. Similar studies examining the relationship between cell volume and the rate of entry into S phase in three established immortal cell lines revealed positive correlation between the rate of entry into S phase and cell volume throughout the size range of the G1 population. This latter observation suggests that the factors involved in the initiation of the S phase may be present in concentrations that are not rate limiting in immortal cell lines.
Pendergrass
W R
WR
Department of Pathology, University of Washington, Seattle 98195.
Angello
J C
JC
Kirschner
M D
MD
Norwood
T H
TH
D002455:Cell Division / Q000502:physiology; D002461:Cell Line, Transformed; D004257:DNA Polymerase II / Q000502:physiology*; D004261:DNA Replication / Q000502:physiology*; D005347:Fibroblasts / Q000166:cytology* / Q000201:enzymology; D005434:Flow Cytometry; D016193:G1 Phase / Q000502:physiology; D006367:HeLa Cells; D006801:Humans; D016196:S Phase / Q000235:genetics / Q000502:physiology
D004257:DNA Polymerase II