Critical nuclear DNA size and distribution associated with S phase initiation. Peripheral location of initiation and termination sites.
Using HeLa S-3 cells synchronized by selective detachment, in this paper we report a parallel study of nuclear morphology and autoradiography grain patterns between middle G1 and middle S phases. Our results show two distinct [3H]-thymidine labeling patterns. The first "peripheral" labeling pattern has a characteristic nuclear size distribution, in contrast to the heterogeneous and varying size distributions of Feulgen-stained nuclei, and apparently is characteristic of very early S phase. The sizes of the second labeling pattern--homogeneous or inhomogeneous grain distribution throughout the nucleus--are equal or larger than the first and vary with S phase progression. Together, the corresponding nuclear sizes of the labeled nuclei represent the larger extreme of nuclear areas, and the labeling index closely parallels the fraction of nuclei with areas larger than the minimum size of the labeled nuclei. These results suggest a characteristic nuclear size (reflecting unique intranuclear DNA distribution) as a necessary, if not sufficient, requirement for S phase initiation. Parallel experimentation with rat liver cells-synchronized in vivo by partial hepatectomy and analyzed by thin section autoradiography--confirms the existence of a peripheral labeling pattern in both the very early part and the very late part of S phase, which reconciles our data with previous results and points to the fact that both initiation and termination sites for DNA replication are near the nuclear periphery.
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Nicolini
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Belmont
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Martelli
D000818:Animals; D001345:Autoradiography; D002455:Cell Division; D002467:Cell Nucleus / Q000502:physiology; D004247:DNA / Q000096:biosynthesis*; D004261:DNA Replication; D006367:HeLa Cells / Q000166:cytology; D006651:Histocytochemistry; D006801:Humans; D008099:Liver / Q000166:cytology / Q000648:ultrastructure; D008297:Male; D010316:Particle Size; D051381:Rats; D011919:Rats, Inbred Strains; D013936:Thymidine / Q000378:metabolism; D014316:Tritium
D014316:Tritium; D004247:DNA; D013936:Thymidine