Preferential inhibition of rDNA transcription by 5-bromodeoxyuridine

1977 ◽  
Vol 25 (1) ◽  
pp. 95-102
Author(s):  
A.E. Lykkesfeldt ◽  
H.A. Andersen

On a chemically defined growth medium the degree of substitution of thymidine with 5-bromodeoxyuridine (BUdR) in DNA of Tetrahymena pyriformis was controlled by the concentration of tetrahydrofiolic acid, BUdR and thymidine in the medium. A correlation between the degree of BUdR substitution in DNA and the reduction in rate of total RNA synthesis has been established. It was found that the reduction of total RNA synthesis results from inhibition of transcription of all RNA species which have been measured. However, independent of the degree of BUdR substitution in DNA, a preferential inhibition of the synthesis of 25s and 17s ribosomal RNA was found. It is concluded that the various genes may respond differently to BUdR substitution with respect to transcription.

1979 ◽  
Vol 35 (1) ◽  
pp. 17-24
Author(s):  
H.A. Andersen ◽  
S.J. Nielsen

The rate of ribosomal RNA synthesis varies greatly with the population density in both exponentially and synchronously growing populations of Tetrahymena pyriformis. Shortly after inoculation of the population - at relatively low cell densities - a gene-dose effect dominates the picture, and a doubling in the gene number is immediately followed by a doubling in the rate of RNA synthesis. However, also other mechanisms are controlling the rate of RNA synthesis. Generally one finds high rates of RNA synthesis in the lag phase of newly inoculated cells, decreasing rate of RNA synthesis during most of the exponential growth phase and very low rate of synthesis in stationary phase cells. We now have results which show that the repression of RNA synthesis in densely populated cultures is caused by a dialysable factor, which is secreted by the cells. If cells are inoculated on a medium which contains this factor the high initial rate of RNA synthesis normally observed is prevented, but the cells multiply and grow with normal generation time until normal stationary-phase population densities are reached.


1978 ◽  
Vol 31 (1) ◽  
pp. 13-23
Author(s):  
J. Keiding ◽  
H.A. Andersen

Ribosomal RNA is synthesized at constant rate during most of the cell cycle in heat-shock synchronized populations of Tetrahymena pyriformis. Early in each macronuclear S-period the rate of synthesis increases abruptly, concomitant with replication of the genes coding for ribosomal RNA. The increase is prevented by inhibitors of DNA replication, added prior to the S-period. Similarly, in cultures synchronized by starvation/refeeding, inhibition of DNA replication, at the time when the rDNA is replicated, will prevent the normal increase in rate of RNA synthesis which follows refeeding. We conclude that inhibition of rDNA replication interferes with the synthesis of rRNA, and we suggest that with respect to rRNA synthesis a gene dosis effect is operating in fast-growing Tetrahymena cells.


1975 ◽  
Vol 17 (3) ◽  
pp. 495-502
Author(s):  
A.E. Lykkesfeldt ◽  
H.A. Andersen

Tetrahymena pyriformis was grown on chemically defined medium in the presence of 5-bromodeoxyuridine (BUdR). Under these growth conditions more than 60% of the thymidine sites in DNA were substituted with BUdR. It was found that RNA synthesis was strongly inhibited by the presence of BUdR in DNA. To assure that incorporation of BUdR into DNA was a prerequisite of the effect observed, BUdR was added to synchronously dividing cells. BUdR had no effect on the cells when present outside the period of nuclear DNA replication, whereas RNA synthesis was strongly inhibited as soon as the genes coding for ribosomal RNA had replicated in the presence of BUdR.


1980 ◽  
Vol 45 (1) ◽  
pp. 31-39
Author(s):  
H.A. Andersen ◽  
A.E. Lykkesfeldt ◽  
S.J. Nielsen

Tetrahymena cells secrete a factor which inhibits RNA synthesis in vivo and in vitro. The factor is a relatively small peptide with a molecular weight between 300 and 1500 Daltons. Other, non-specific peptides in the broth medium or added to a chemically defined medium have a stimulatory effect on RNA synthesis in vivo and such peptides also stimulate the in vitro synthesis of RNA in a r-chromatin preparation. On the basis of these results we conclude that such extracellular small peptides compete with a specific factor which is part of the intracellular regulatory mechanism controlling the rate of RNA synthesis. The consequence of such competition is a high overproduction of ribosomal RNA in cells inoculated on peptide-rich broth media.


1975 ◽  
Vol 78 (2) ◽  
pp. 401-416 ◽  
Author(s):  
Anja Isotalo ◽  
R. S. Santti

ABSTRACT The total RNA isolated at various times up to 24 h after testosterone administration from the ventral prostate of castrated rats, was labelled either by injecting 3H-orotic acid directly into the ventral prostate 6 h before the animals were killed, or by incubating prostatic tissue in vitro with 3H-uridine for 20 to 60 min. The isolated RNA was separated into tRNA, ribosomal RNA (Q1 RNA) and two DNA-like RNA fractions (Q2) and TD RNA) by chromatography on methylated albumin kieselguhr (MAK) columns, and the fractions were further analysed by sucrose gradient centrifugation. Testosterone given into castrated animals for 12 h, stimulated the labelling of all main fractions. The radioactivity of TD RNA after a 60-min incubation period in vitro with 3H-uridine was approximately twice that seen in the castrated rat, while there was a 3.1- and 2.2-fold increase in the radioactivity of the Q1 and Q2 RNA fractions respectively. Kinetics of incorporation of 3H-uridine into different RNA fractions revealed that the hormone facilitated the labelling of the TD RNA fraction relatively more than that of the Q2 fraction. The injection of 3H-orotic acid into the ventral prostate labelled the Q1 RNA preferentially. More than 60 % of the recovered radioactivity was found in Q1 RNA (as 18 and 28 S). Testosterone increased markedly (9.4-fold) the labelling of this fraction. It was concluded that testosterone has an activatory effect on the production of ribosomal RNA, and the bulk of the testosterone effect on the total RNA labelling is to be found in this fraction. Furthermore, it seems likely that testosterone also stimulates both the synthesis and processing of DNA-like RNA. When actinomycin D was given 2 h before the hormone administration in a dose of 25 μg per 100 g of body weight, there was no noticeable increase in the labelling of any fraction above the level seen in the untreated castrated rat. There is evidence that testosterone exerts some effects on the labelling of proteins with radioactive amino acids and 14C-glucose metabolism in the absence of that fraction of the total RNA synthesis which is sensitive to a low dose (25 μg per 100 g of body weight) of actinomycin D (Isotalo & Santti 1972). In this way it may be concluded that the major changes of the RNA synthesis after testosterone administration are likely to be secondary to the protein synthesis and glucose metabolism, or the hormone exerts its anabolic effect on prostatic cells at different sites and by different modes of action, each of which can be operated independently.


1989 ◽  
Vol 264 (30) ◽  
pp. 18220-18227
Author(s):  
P J McDermott ◽  
L I Rothblum ◽  
S D Smith ◽  
H E Morgan

1986 ◽  
Vol 261 (11) ◽  
pp. 5187-5193
Author(s):  
M N Schnare ◽  
T Y Heinonen ◽  
P G Young ◽  
M W Gray

Genetics ◽  
1973 ◽  
Vol 73 (3) ◽  
pp. 429-434
Author(s):  
J James Donady ◽  
R L Seecof ◽  
M A Fox

ABSTRACT Drosophila melanogaster embryos that lacked ribosomal DNA were obtained from appropriate crosses. Cells were taken from such embryos before overt differentiation took place and were cultured in vitro. These cells differentiated into neurons and myocytes with the same success as did wild-type controls. Therefore, ribosomal RNA synthesis is not necessary for the differentiation of neurons and myocytes in vitro.


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