scholarly journals Template activity of chromatin during stimulation of cellular proliferation in human diploid fibroblasts

1971 ◽  
Vol 122 (2) ◽  
pp. 189-195 ◽  
Author(s):  
John Farber ◽  
Giovanni Rovera ◽  
Renato Baserga

1. Contact-inhibited confluent monolayers of WI-38 human diploid fibroblasts can be stimulated to divide by replacing the medium with fresh medium containing 30′ foetal calf serum. 2. Of the cells 40–75′ are stimulated to divide with a peak DNA synthesis between 15 and 21h and a peak mitotic index between 28 and 30h after stimulation. 3. In the first 12h before the initiation of DNA synthesis there is a biphasic increase in the incorporation of [3H]uridine into RNA of whole cells. 4. This is paralleled by a similar biphasic stimulation of chromatin template activity measured in vitro in a system in which purified cell chromatin is incubated with an exogenous RNA polymerase isolated from Escherichia coli. 5. The changes in chromatin template activity are believed to represent activation of the genome, with more sites available for RNA synthesis, and to account almost entirely for the changes in RNA synthesis occurring in the whole cell.

1986 ◽  
Vol 237 (3) ◽  
pp. 741-747 ◽  
Author(s):  
D J McQuillan ◽  
C J Handley ◽  
H C Robinson

Proteoglycan synthesis in explant cultures of adult bovine articular cartilage is stimulated in a dose-dependent manner when the tissue is cultured in the presence of foetal-calf serum. The stimulation of proteoglycan synthesis is paralleled by a similar increase in DNA synthesis; however, when DNA synthesis is inhibited by hydroxyurea the stimulation of proteoglycan synthesis by serum remains essentially the same. The apparent half-life of the pool of proteoglycan core protein precursor was measured in freshly isolated tissue as well as in tissue cultured for 7 days in the presence and in the absence of foetal-calf serum; under all conditions the half-life was the same, suggesting that this value is independent of the net rate of proteoglycan synthesis. In the presence of actinomycin D, an inhibitor of RNA synthesis, there was a difference in the apparent half-life of the available pool of mRNA coding for proteoglycan core protein: 8.5 h for tissue maintained in the presence of serum and 3.8 h for tissue cultured in the absence of serum. It is suggested that proteoglycan synthesis is stimulated by serum factors at the level of DNA-dependent RNA synthesis. Concomitant with an increase in the rate of proteoglycan synthesis induced by the presence of serum in the culture medium, an increase in the concentrations of several glycosyltransferases involved in chondroitin sulphate synthesis was also observed.


1971 ◽  
Vol 134 (5) ◽  
pp. 1095-1113 ◽  
Author(s):  
M. R. Pandian ◽  
G. P. Talwar

The effect of pituitary growth hormone on the biosynthesis of DNA in the thymus and other lymphoid organs, as well as the ability of the rat to respond immunologically to sheep red blood cells, has been evaluated. There is a marked reduction in plaque-forming cells, hemagglutination titers, and DNA synthesis in animals when examined at 15 wk after hypophysectomy. Administration of bovine growth hormone (BGH) leads to the enhancement of DNA synthesis in lymphoid organs and recovery of the immune response. Similar effects of the hormone are observed in plateaued rats. Injection of rabbit anti-BGH globulins, in contrast to normal rabbit globulins, over 5 days causes a drop in the weight of the thymus and in the rate of DNA synthesis in this organ. The thymus is also the organ in which stimulation of DNA synthesis is observed at a time period earlier than the spleen and lymph nodes after a single injection of BGH. The hormone stimulates not only the incorporation of thymidine-3H into DNA in the cortical cells, but also the incorporation of sodium sulfate-35S into TCA-insoluble biopolymers reported to be elaborated in the medullary area of the thymus. An in vitro system for the action of BGH on the thymus has been described. There is an obligatory requirement for calcium, but not for fetal calf serum in the medium for the hormone effect. An early action of the hormone is the enhanced incorporation of uridine-G-3H into RNA in thymocytes which is followed by a stimulation of the synthesis of proteins and DNA. The stimulatory action of growth hormone on RNA synthesis is not because of a facilitated uptake of the radioactive uridine by the cells under hormonal influence, a mechanism by which insulin is observed to increase RNA synthesis in thymocytes in vitro. The action of growth hormone on thymocytes is specific, since thyroid-stimulating hormone (TSH), luteinizing hormone (LH), and heat-inactivated growth hormone are not effective. BGH has also a beneficial action on the regeneration of the thymus and spleen in starved rats.


Virology ◽  
1979 ◽  
Vol 95 (1) ◽  
pp. 222-226 ◽  
Author(s):  
Tohru Kamata ◽  
Shigeaki Tanaka ◽  
Shogo Aikawa ◽  
Yorio Hinuma ◽  
Yasushi Watanabe

1977 ◽  
Vol 63 (1) ◽  
pp. 31-41 ◽  
Author(s):  
Rosanna Supino ◽  
Anna M. Casazza ◽  
Aurelio Di Marco

This paper reports the effects of daunorubicin and adriamycin on DNA and RNA synthesis of in vitro cultured mouse embryo fibroblasts (MEF) stimulated by fetal calf serum (FCS). The addition of FCS to quiescent MEF cultures brings about a wave of RNA synthesis, followed by DNA synthesis which starts between 8 and 12 h after change of medium and proceed for up to 24 h. These cells are therefore partially synchronized. The level of DNA synthesis depends on the amount of FCS added. Daunorubicin and adriamycin are almost equally effective in inhibiting DNA synthesis, as well as cell proliferation, which takes place later. Adriamycin is more active than daunorubicin on RNA synthesis. In cultures treated for an 8 h period starting at different times after FCS addition, the highest DNA synthesis inhibition is achieved by treatment during the first 8 h, when DNA synthesis has not yet started. The cellular uptake of daunorubicin is constantly higher than that of adriamycin, in any experimental condition tested. The results show that FCS-stimulated MEF can provide a simple method for studying the effects of anthracycline antibiotics on partially synchronized cells.


1980 ◽  
Vol 105 (3) ◽  
pp. 503-512 ◽  
Author(s):  
Terry I. Korotzer ◽  
James A. Clagett ◽  
William P. Kolb ◽  
Roy C. Page

1982 ◽  
Vol 50 (1) ◽  
pp. 11-15
Author(s):  
B. De Jong ◽  
G.J.P.A. Anders ◽  
I.H. Van Der Meer ◽  
J. Zijlstra ◽  
V.J.S. Idenburg

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