Changes in template activity and structure of nuclei from WI-38 cells in the prereplicative phase.
Quiescent confluent monolayers of WI-38 fibroblasts were stimulated to proliferate by either adding 10% fetal calf serum or by trypsinization and replating at lower density. The length of the prereplicative phase was 12 hr after serum stimulation and 18 hr after trypsinization and replating at lower density. Nuclei were isolated from WI-38 cells at different time intervals after either type of stimulation and their template activity, circular dichroism spectra, and ability to bind ethidium bromide were investigated. All these parameters were similarly increased after either type of stimulation. However, these changes, like the onset of DNA synthesis, were delayed 6 hr in cells trypsinized and replated at lower density. While there were no detectable changes in nuclear protein content after serum stimulation, at least 40% of nuclear protein, mostly nonhistone chromosomal proteins, were lost after trypsinization. The amount of nuclear proteins returned to prestimulation levels only 6-8 hr after replating. These data seem to suggest that nonhistone chromosomal proteins lost by trypsinization are essential for the entrance of WI-38 cells into the "prereplicative phase".
D000818:Animals; D001665:Binding Sites; D002417:Cattle; D002455:Cell Division; D002460:Cell Line; D002467:Cell Nucleus / Q000378:metabolism* / Q000648:ultrastructure; D002942:Circular Dichroism; D003470:Culture Media; D004261:DNA Replication; D004996:Ethidium; D008168:Lung / Q000196:embryology; D009690:Nucleic Acid Conformation; D013056:Spectrophotometry, Ultraviolet; D013698:Templates, Genetic; D013936:Thymidine / Q000378:metabolism; D013997:Time Factors; D014357:Trypsin
D003470:Culture Media; D014357:Trypsin; D004996:Ethidium; D013936:Thymidine