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Biomedical subjects

R Baserga

Publications and source records attributed to R Baserga.

At least 289 records · Page 16Linked to original sources
PubMed1976

The role of nuclei and nucleoli in the control of cell proliferation.

A number of changes have been reported to occur in chromatin of quiescent cells stimulated to proliferate. These changes, indicative of increased transcriptional activity, occur in the early prereplicative phase, several hours before the onset of DNA replication, and are detectable not only in chromatin, but also in isolated nuclei and in whole cells. Most of the increased transcriptional activity can be attributed to an increased activity of the nucleolus; however, extranucleolar genes are also important in the regulation of the cell cycle flow, from G0 to S.

Animals↗
PubMed1975

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".

Animals↗
PubMed1975

Chromatin changes during the cell cycle of HeHa cells.

HeLa S3 cells were synchronized by selective mitotic detachment and chromatin was isolated from synchronized populations at various times during the cell cycle. The isolated chromatins were studied by circular dichroism and for their ability to bind the intercalating dye, ethidium bromide. Chromatin from cells in the middle of the S phase showed an increase in maximum ellipticity in the 250- to 300-nm region of circular dichroism spectra and an increased ability to bind ethidium bromide, when compared to chromatin isolated from mitotic cells. Chromatin from G1 cells had values intermediate between mitotic and mid-S chromatins.

Binding Sites↗
PubMed1975

Effect of cell trypsinization on nuclear proteins of WI-38 fibroblasts in culture.

When resting confluent monolayers of WI-38 fibrolasts are trypsinized and replated at a lower density they are stimulated to proliferate again with an interval of 18 hours between replating and the onset of DNA synthesis. Trypsinization of resting cells causes a 40% loss of nuclear proteins as well as of cytoplasmic proteins. The amount of nuclear proteins remains low for the first six hours after the cells have been replated and then it increases rapidly, reaching the same level of non-trypsinized resting cells by ten hours after plating. The proteins that are lost from the nucleus immediately after trypsinization are chromatin-associated proteins and most of them are non-histone chromosomal proteins, although a modest loss of histones cannot be ruled out. The loss of non-histone chromosomal proteins from cells that have been trypsinized causes changes in the structure of chromatin that can be detected by circular dichroism and by viscosity measurements. These results show that cell trypsinization causes an extensive loss of proteins from chromatin and that the loss is restored only several hours after the cells have been replated at a lower density.

Cell Line↗
PubMed1975

Circular dichroism and ethidium bromide binding studies of chromatin from WI-38 fibroblasts stimulated to proliferate.

Confluent monolayers of WI-38 diploid fibroblasts can be stimulated to proliferate by fresh serum. In the first 3 h after stimulation (that is, several hours before DNA replication) the chromatin of stimulated cells show structrual changes which include: (1) an increase in maximum positive ellipticity and a blue shift in the 250-300 nm region of circular dichroism spectra; and (2) an increase,in isolated chromatin, of the number of binding sites for the intercalating dye, ethidium bromide.The differences between chromtin of stimulated and chromatin of unstimulated cells are abolised when bother chromatins are treated with 0.25 M NaCL.

Binding Sites↗
PubMed1975

Effect of 5-bromodeoxyuridine on the transcriptional properties of the genome in WI-38 human diploid fibroblasts.

Growth of WI-38 diploid fibroblasts in a medium containing 5-bromodeoxyuridine (BrdU) resulted in an increased GMP and a decreased AMP incorporation into the RNA synthesised in vitro on a chromatin template. This effect was similar to that previously reported using 3T6 mouse fibroblasts-1. Substitution of thymidine by BrdU in DNA, also altered the characteristics of the DNA template itself, since the increased incorporation of guanine and decreased incorporation of adenine into RNA were evident also when purified, isolated DNA was used as template. The extent of replacement of AMP by GMP was proportional to the extent of replacement of thymidine by BrdU. Although there are variations in the base composition of RNA transcribed from BrdU-containing DNA templates, there are no significant difference in overall template activity or in the number of available chromatin binding sites for E. coli RNA polymerase. Confluent monolayers of BrdU-treated WI-38 fibroblasts are still able to respond with cell proliferation to a change of medium, as evidenced by an increased incorporation of (-3H)thymidine and an increase in chromatin template activity. The length of the prereplicative phase is similar in both BrdU-treated and untreated cells, although the magnitude of the increase of (-3H)thymidine incorporation is reduced by approximately 30% after BrdU treatment. The increase in chromatin template activity is associated with an increase in the number of chromatin binding sites for E. COLI RNA polymerase, suggesting that the presence of BrdU does not interfere with the availability of initiation sites or alter the actual rate of transcription.

Adenosine Monophosphate↗
PubMed1975

Effect of chromosomal proteins extractable with low concentrations of NaCl on chromatin structure of resting and proliferating cells.

When quiescent WI-38 cells are stimulated to proliferate by nutritional changes, the chromatin of stimulated cells shows an increased ellipticity in circular dichroism spectra and an increased ability to bind ethidium bromide. The differences between the chromatins of quiescent and stimulated cells are abolished if both chromatins are treated with 0.25 M NaCl. However, when salt-treated chromatins are reconstituted by dialysis agains 10 mM Tris-HCl, the difference in circular dichroism spectra and ethidium bromide binding re-appear. Furthermore, addition of the 0.25 M NaCl extract from chromatin of stimulated cells to chromatin of quiescent cells causes an increased ellipticity in circular dichroism spectra similar to that of stimulated chromatin. Since only proteins (mostly nonhistones) are detectable in the 0.25 M NaCl extract, these results strongly suggest that the chromosomal proteins extractable with low concentrations of salt are responsible for the functional and structural changes occurring in chromatin of WI-38 cells stimulated to proliferate.

Binding Sites↗