共查询到20条相似文献,搜索用时 15 毫秒
1.
Analysis of the biological effects of specific DNA alkylations by simple alkylating agents is complicated by the variety of sites involved. It is, therefore, of value to be able to incorporate into cellular DNA nucleosides alkylated in a single position, e.g., O6-methyldeoxyguanosine. Such cellular incorporation is particularly difficult to achieve because this nucleoside is rapidly demethylated by adenosine deaminase. We have attempted to achieve such incorporation into the DNA of V79 cells by using coformycin, an inhibitor of adenosine deaminase, and by forcing the cells to depend on exogenous purines by the use of medium containing aminopterin. The DNA of V79 cells exposed to O6-methyl-[8-3H]deoxyguanosine (2.4 μM, sp. act. 14 500 Ci/mole) showed an incorporation level of 4 × 10−8 nucleotides. When 1000-fold higher concentrations were employed (3–15 mM, sp. act. 1.6 Ci/mole), significant cytotoxicity and inhibition of DNA synthesis was observed. However, because it was not economically feasible to administer high specific activity O6-methyldeoxyguanosine to the cells at these concentrations, we could not determine the amount of labeled nucleoside incorporated into DNA. Examination of the frequency of 6-thioguanine-resistant cells in these treated populations showed no significant increase above the background level. Comparison of the cytotoxic effect of O6-methyldeoxyguanosine with deoxyadenosine showed that the toxicity induced by O6-methyldeoxyguanosine could have resulted from mimicry of deoxyadenosine, rather than by incorporation of the alkylated nucleoside itself. 相似文献
2.
Antibodies to O6-methyldeoxyguanosine were produced in rabbits and utilized in a radioimmunoassay to detect this nucleoside at picomole levels. The specificity of the antibodies was demonstrated by the use of nucleoside analogues as inhibitors in the radioimmunoassay. The antibodies cross-reacted with O6-methylguanosine, O6-methylguanine, and O6-ethylguanosine. There was 10(4) to 10(6) times less sensitivity to inhibition by deoxyadenosine, deoxyguanosine, and guanosine than by O6-methyldeoxyguanosine. The radioimmunoassay also detected O6-methylguanine in DNA alkylated by agents known to produce O6-methylguanine, such as N'-methyl-N-nitrosourea. DNA alkylated with dimethyl sulfate, which does not produce O6-methylguanine in DNA, cross-reacted with the antibodies to a very limited extent. Such an assay system for modified nucleic acid components would be very useful in following the production, persistence, and repair of these lesions in a variety of cells and tissues treated with a broad spectrum of carcinogens and suspected carcinogens. 相似文献
3.
We have shown previously that a low concentration of tritiated deoxyadenosine, i.e., 1 µCi/ml, selectively kills wild-type S49 murine lymphoma cells. Mutant cells resistant to [3H]deoxyadenosine lacked adenosine kinase completely but retained a significant level of deoxyadenosine phosphorylating activity. To study further the specificity of [3H]deoxyadenosine selection, lymphoma cell clones resistant to 15 µCi/ml [3H]deoxyadenosine have been derived. The resistant line, S49-dA15, is also resistant to high levels of nonradioactive deoxyadenosine and to deoxyguanosine but remains sensitive to thymidine. The thymidine inhibition of the growth of the mutant, in contrast to that of the wild-type cells, cannot be prevented by deoxycytidine. The mutant line lacks deoxycytidine kinase that also phosphorylates deoxyadenosine. In addition, the mutant cells excrete a large amount of deoxycytidine into culture medium, consistent with a failure of salvage of the nucleoside in the absence of an appropriate kinase, i.e., deoxycytidine kinase. In contrast, a deoxycytidine kinase-deficient cell line that was selected with arabinosylcytosine does not excrete deoxycytidine and contains high deoxycytidine deaminase activity. [3H]Deoxyadenosine can be used as a selective agent for specific selection of deoxycytidine kinase-negative mutants. 相似文献
4.
The pro-mutagenicity of chemically-induced methylation of DNA at the O6 position of dexoyguanosine was studied in cultured adult rat liver epithelial cells. To modify the level of O6-methyldeoxyguanosine (O6-medGuo) resulting from exposure to an alkylating agent, partial depletion of the O6-alkylguanine-DNA alkyltransferase (AGT) repair system was produced by pretreatment of ARL 18 cells with a non-toxic dose of exogenous O6-methylguanine (O6-meG). Exposure of cells to 0.6 mM O6-meG for 4 h depleted AGT activity by about 40%. Intact and pretreated cells were exposed to a range of doses of N-methyl-N′-nitro-N-nitrosoguanidine (MNNG), and mutagenesis at the hypoxanthine-guanine phosphoribosyl transferase (HGPRT) locus was quantified by measurement of 6-thioguanine-resistant mutants. The mutagenicity of MNNG was dose dependent and was greater in O6-meG pretreated cultures than in intact cultures. Immunoslot blot measurement of O6-medGuo employing a mouse monoclonal antibody demonstrated that MNNG produced O[su6-medGuo and that the intact liver cells were efficient in eliminating this lesion from their DNA. Since depletion of AGT would be expected to affect the rate of elimination of only O6-medGuo, it is concluded that this lesion is highly pro-mutagenic. 相似文献
5.
Lyle A. Dethlefsen 《Cell proliferation》1974,7(3):213-222
The toxicity of 3H-5-iodo-2′-deoxyuridine (3H-IUdR) was evaluated by injecting tumor-bearing C3H mice with different concentrations of ethanol (the solvent), different doses of tritium tagged onto either IUdR or thymidine and different chemical doses of IUdR, and then measuring the 3H-IUdR incorporation into duodenal and mammary tumor DNA as well as the cellular kinetics of duodenal crypt cells. Ethanol (37% or less, 0.2 ml/mouse) does not significantly inhibit IUdR incorporation into DNA, and the incorporation after a tritium dose of 75 μCi 3H-IUdR/mouse (about 3 μCi/g body weight) is not less than the incorporation following an injection of 25 μCi 3H-IUdR/mouse when the IUdR dose is below 0.005 μmole per mouse. The toxic effects are primarily due to chemical toxicity from IUdR per se. IUdR, at doses of 0.2 μmoles per mouse does inhibit IUdR incorporation into duodenal and tumor DNA, and the duodenal labeling index and the fraction of labeled mitoses are significantly reduced when 0.013 μmole IUdR per mouse is injected. Also some of the duodenal cells containing IUdR apparently undergo only one post-labeling division and the generation time (Tc) of the cells containing IUdR (25 μCi 3H-IUdR/mouse) is 15.3 hr as compared to 13.3 hr for cells labeled with 3H-T (75 μCi/mouse). This increase in Tc is probably not statistically significant; nevertheless, these results do indicate that one must be exceedingly cautious when using 3H-IUdR as a radiotracer for studies concerned with in vivo cellular kinetics and, at least for C3H mice, the dose should be less than 0.01 μmole per 25 g mouse. 相似文献
6.
Takats ST 《Plant physiology》1967,42(7):907-910
The uptake of 3H-thymidine into pollen grains of Tradescantia paludosa was studied in the presence of 2′-deoxyadenosine. 1) Millimolar deoxyadenosine caused an immediate inhibition of incorporation of 3H-thymidine into DNA extracted with hot trichloroacetic acid. 2) The radioactivity in acid-soluble derivatives of 3H-thymidine was examined by paper chromatography and, following incubation of pollen grains in the presence of millimolar deoxyadenosine, was found to be increased several-fold in 3H-deoxythymidine triphosphate. 3) The time-course of inhibition showed that the acid-soluble derivatives of 3H-thymidine accumulated initially at a rate unaffected by deoxyadenosine, despite the nearly complete inhibition of incorporation of 3H-thymidine into DNA. This is discussed in relation to possible mechanisms of inhibition by deoxyadenosine. 相似文献
7.
The alkylating antitumor agent triethyleneiminobenzoquinone (Trenimon) causes a rapid decrease in the incorporation of labeled thymidine into the DNA of Yoshida or Ehrlich ascites tumor cells. The effect is expressed 4 h after administration of 6 × 10−8 moles/kg of the drug to mice bearing Yoshida ascites tumors or of 6 × 10−7 moles/kg to Ehrlich ascites tumor-bearing animals, respectively. The reduced incorporation of labeled thymidine which is observed under these conditions is not due to an inhibition of DNA synthesis. DNA synthesis was measured by an isotope dilution assay after pulse-labeling with 3H-thymidine and by monitoring the increase in the total amount of DNA of the cell populations. The data demonstrate that DNA synthesis is not affected during the first 8 h after exposure to the drug. This conclusion is supported by cell kinetic measurements which indicate that the alkylating agent does not interfere with the progression of cells into the S phase, but exerts a block at the G 2 stage of the cell cycle. The reduced incorporation of thymidine into DNA is explained by a decreased transport of the nucleoside into the cells. 相似文献
8.
Gerda Tyrsted 《Experimental cell research》1975,91(2)
Requirements and optimal conditions have been studied for measurements of dGTP and dCTP in cellular extracts using the copolymer [d(1 − C)] as primer in a reaction catalysed by the large fragment of DNA polymerase from E. coli. The pool size of dGTP and dCTP in the human lymphocytes in the absence of PHA was found to be about 0.1 and 0.15 pmoles/106 cells, respectively. After treatment with PHA the pool size of both deoxynucleotides increased. The pool size of dCTP reached a maximum after 67 h simultaneously with the peak value of labelled deoxythymidine incorporation into DNA and the variation in these two parameters was very similar. The variation in the dGTP pool, however, was not so distinctly related to deoxythymidine incorporation as in the dCTP pool, since the increase in the dGTP pool was very small from 52–67 h. During transformation the dGTP pool was found to be the smallest pool. The relative cellular content of mono-, di- and triphosphate esters of deoxyadenosine, deoxyguanosine and deoxycytidine was studied. 相似文献
9.
A chromatographic procedure for improved separation of deoxyribonucleosides and methylated deoxyribonucleosides is described. DNA was isolated from liver and small intestine of rats treated with [14C]dimethylnitrosamine ([14C]DMN) or ([3H]MNU), and the purified DNA was hydrolyzed enzymatically. The deoxyribonucleosides were chromatographed on an Aminex A-6 cation exchange column at 37°C with 0.4 M ammonium formate, pH 4.5, as eluant. In addition to showing the presence of the expected alkylated products, N7-methyldeoxyguanosine (determined as N7-methylguanine) and O6-methyldeoxyguanosine, several other minor methylated products were found in liver and intestinal DNA of rats treated with DMN or MNU. Two of these products are believed to be N3-methylthymidine and O4-methylthymidine. 相似文献
10.
Covalent capture of a human O(6)-alkylguanine alkyltransferase-DNA complex using N(1),O(6)-ethanoxanthosine, a mechanism-based crosslinker
下载免费PDF全文
![点击此处可从《Nucleic acids research》网站下载免费的PDF全文](/ch/ext_images/free.gif)
The DNA repair protein O6-alkylguanine alkyltransferase (AGT) is responsible for removing promutagenic alkyl lesions from exocyclic oxygens located in the major groove of DNA, i.e. the O6 and O4 positions of guanine and thymine. The protein carries out this repair reaction by transferring the alkyl group to an active site cysteine and in doing so directly repairs the premutagenic lesion in a reaction that inactivates the protein. In order to trap a covalent AGT–DNA complex, oligodeoxyribonucleotides containing the novel nucleoside N1,O6-ethanoxanthosine (eX) have been prepared. The eX nucleoside was prepared by deamination of 3′,5′-protected O6-hydroxyethyl-2′-deoxyguanosine followed by cyclization to produce 3′,5′-protected N1,O6-ethano-2′-deoxyxanthosine, which was converted to the nucleoside phosphoramidite and used in the preparation of oligodeoxyribonucleotides. Incubation of human AGT with a DNA duplex containing eX resulted in the formation of a covalent protein–DNA complex. Formation of this complex was dependent on both active human AGT and eX and could be prevented by chemical inactivation of the AGT with O6-benzylguanine. The crosslinking of AGT to DNA using eX occurs with high yield and the resulting complex appears to be well suited for further biochemical and biophysical characterization. 相似文献
11.
《Nucleosides, nucleotides & nucleic acids》2013,32(12):1849-1864
The formation of a disaccharide nucleoside (11) by O3′‐glycosylation of 5′‐O‐protected 2′‐deoxyadenosine or its N 6‐benzoylated derivative has been observed to be accompanied by anomerisation to the corresponding α‐anomeric product (12). The latter reaction can be explained by instability of the N‐glycosidic bond of purine 2′‐deoxynucleosides in the presence of Lewis acids. An independent study on the anomerisation of partly blocked 2′‐deoxyadenosine has been carried out. Additionally, transglycosylation has been utilized in the synthesis of 3′‐O‐β‐d‐ribofuranosyl‐2′‐deoxyadenosines and its α‐anomer. 相似文献
12.
F. Wanka 《Experimental cell research》1974,85(2)
The effect of a pre-treatment with 2 mM deoxyadenosine on DNA synthesis in bovine liver cells in vitro was investigated. The incorporation of 3H-deoxythymidine was strongly depressed after 1 h treatment and recovered only gradually during the following 4 to 6 h. Addition of 50 μM deoxyguanosine during this time effectively counteracted the depression. An autoradiographic study revealed that the depression was due to a decrease in thymidine incorporation per cell rather than a decrease of the proportion of thymidine-incorporating cells. It is concluded from these results that the rate of DNA synthesis is decreased during the initial hours after removal of the deoxy-adenosine because of a continued deficiency of dGTP. This conclusion is supported by the appearance of a high proportion of 3H-label in replicating intermediates after 1 h of incorporation which is otherwise only observed after pulse labelling for 5 to 10 min. 相似文献
13.
To find general metabolic profiles of purine ribo- and deoxyribonucleotides in potato (Solanum tuberosum L.) plants, we looked at the in situ metabolic fate of various 14C-labelled precursors in disks from growing potato tubers. The activities of key enzymes in potato tuber extracts were also studied. Of the precursors for the intermediates in de novo purine biosynthesis, [14C]formate, [2-14C]glycine and [2-14C]5-aminoimidazole-4-carboxyamide ribonucleoside were metabolised to purine nucleotides and were incorporated into nucleic acids. The rates of uptake of purine ribo- and deoxyribonucleosides by the disks were in the following order: deoxyadenosine > adenosine > adenine > guanine > guanosine > deoxyguanosine > inosine > hypoxanthine > xanthine > xanthosine. The purine ribonucleosides, adenosine and guanosine, were salvaged exclusively to nucleotides, by adenosine kinase (EC 2.7.1.20) and inosine/guanosine kinase (EC 2.7.1.73) and non-specific nucleoside phosphotransferase (EC 2.7.1.77). Inosine was also salvaged by inosine/guanosine kinase, but to a lesser extent. In contrast, no xanthosine was salvaged. Deoxyadenosine and deoxyguanosine, was efficiently salvaged by deoxyadenosine kinase (EC 2.7.1.76) and deoxyguanosine kinase (EC 2.7.1.113) and/or non-specific nucleoside phosphotransferase (EC 2.7.1.77). Of the purine bases, adenine, guanine and hypoxanthine but not xanthine were salvaged for nucleotide synthesis. Since purine nucleoside phosphorylase (EC 2.4.2.1) activity was not detected, adenine phosphoribosyltransferase (EC 2.4.2.7) and hypoxanthine/guanine phosphoribosyltransferase (EC 2.4.2.8) seem to play the major role in salvage of adenine, guanine and hypoxanthine. Xanthine was catabolised by the oxidative purine degradation pathway via allantoin. Activity of the purine-metabolising enzymes observed in other organisms, such as purine nucleoside phosphorylase (EC 2.4.2.1), xanthine phosphoribosyltransferase (EC 2.4.2.22), adenine deaminase (EC 3.5.4.2), adenosine deaminase (EC 3.5.4.4) and guanine deaminase (EC 3.5.4.3), were not detected in potato tuber extracts. These results suggest that the major catabolic pathways of adenine and guanine nucleotides are AMP → IMP → inosine → hypoxanthine → xanthine and GMP → guanosine → xanthosine → xanthine pathways, respectively. Catabolites before xanthosine and xanthine can be utilised in salvage pathways for nucleotide biosynthesis. 相似文献
14.
《Nucleosides, nucleotides & nucleic acids》2013,32(5-8):635-639
Abstract A facile synthesis of oligodeoxynucleotides (ODN) containing 2′-deoxy-6-thioinosine (dI6S) based on the convertible nucleoside O6-phenyl-2′-deoxyinosine is presented. After standard solid-phase DNA synthesis and removal of the cyanoethyl protecting groups with DBU treatment with aqueous sodium hydrogen sulfide introduces the sulfur functionality, deprotects the other nucleobases and cleaves the ODN from the solid support in a one-pot reaction. In addition, the extinction coefficient of 2′-deoxy-6-thioinosine is determined by enzymatic fragmentation of the resulting ODN in the presence of adenosine deaminase. 相似文献
15.
《Biochemical medicine》1981,25(3):288-297
The apparent rate of phosphorylation of deoxyadenosine has been studied in crude extracts of human leukemic cells. Since detection and quantitation of phosphorylated products of deoxyadenosine are not possible in the presence of the competing enzyme, adenosine deaminase, an assay system has been devised in which deaminase activity is totally inhibited. Two inhibitors of adenosine deaminase, erythro-9-(2-hydroxy-3-nonyl)adenine and 2′-deoxycoformycin were tested. Complete inhibition of adenosine deaminase cannot be achieved with erythro-9-(2-hydroxy-3-nonyl)adenine, but can be achieved with 2′-deoxycoformycin at concentrations greater than 100 μ m. Use of deoxycoformycin allows an accurate assessment of phosphorylation of deoxyadenosine and its nucleoside analogs. Cellular extracts from patients with several types of leukemia contained a 30-fold difference in relative deoxyadenosine kinase activity. Adenine arabinoside is a competitive inhibitor of deoxyadenosine, but not adenosine phosphorylation with a Ki (app) of 3.2 mm. This inhibition pattern is consistent with a common pathway of phosphorylation for deoxyadenosine and adenine arabinoside in human leukemic cells. 相似文献
16.
Specifically alkylated DNA fragments. Synthesis and physical characterization of d[CGC(O6Me)GCG] and d[CGT(O6Me)GCG] 总被引:5,自引:1,他引:4
下载免费PDF全文
![点击此处可从《Nucleic acids research》网站下载免费的PDF全文](/ch/ext_images/free.gif)
Two hexamer DNA fragments containing a carcinogenic modified base, O6-methyl guanine, have been synthesized by a solid-phase phosphotriester method, in which the unmodified guanine residues present were O6 protected with the 4-nitrophenylethyl group. These two alkylated oligonucleotides were found to have similar Tm's about 40° lower than the unmodified parent compund, d(CG)3. Moreover, the presence of the (O6Me)G appears to inhibit the B→Z transition, as determined by CD spectroscopy. 相似文献
17.
Intracellular conversions of deoxyribonucleosides by Novikoff rat hepatoma cells and effects of hydroxyurea 总被引:3,自引:0,他引:3
The incorporation of 3H-labeled deoxyadenosine and deoxyguanosine into nucleic acids by cultured Novikoff rat hepatoma cells is about 80% into RNA and 20% into DNA. The pathways of incorporation have been elucidated in studies with whole cells and cell-free extracts. Deoxyadenosine is very rapidly deaminated to deoxyinosine. Most of the deoxyinosine formed by whole cells is transported out of the cells and accumulates in the medium. A portion of the deoxyinosine, and deoxyguanosine are phosphorolyzed by purine nucleoside phosphorylase to hypoxanthine and guanine, respectively. The latter are subsequently converted by hypoxanthine-guanine phosphoribosyl transferase to IMP and GMP, respectively. Incorporation of the purine deoxyribonucleosides into DNA is mainly via this pathway and the subsequent reduction of ADP and GDP by ribonucleoside reductase, although a small proportion of the deoxyadenosine and deoxyguanosine taken up by the cells seems to be directly phosphorylated to dAMP and dGMP, respectively. Deoxyguanosine is incorporated only into guanine residues of RNA and DNA. Deoxyadenosine is also mainly incorporated into guanine residues of RNA and DNA, although the radioactivity of deoxyadenosine in the acid-soluble pool is almost exclusively associated with ATP. A similar labeling pattern is observed with labeled deoxyinosine, inosine or hypoxanthine. The pyrimidine deoxyribonucleosides, on the other hand, are specific precursors for their respective bases in DNA. Hydroxyurea inhibits the incorporation of all deoxyribonucleosides into DNA. Results from pulse-chase experiments indicate that the inhibition of DNA synthesis is prevented by the presence of high concentrations of deoxyadenosine plus deoxyguanosine in the medium. Either purine deoxyribonucleoside alone or deoxycytidine, hypoxanthine or inosine alone or in combination with deoxyadenosine or deoxyguanosine are ineffective. The results are consistent with the conclusion that the inhibition of DNA synthesis is due to a depletion of the dATP and dGTP pools as a result of the hydroxyurea treatment. On the other hand, hydroxyurea causes an increased incorporation of thymidine and deoxycytidine into the dTTP and dCTP pools, respectively. Evidence is presented to indicate that this effect of hydroxyurea is due to an increased synthesis of dTTP and dCTP rather than to an inhibition of their turnover. 相似文献
18.
The high susceptibility of certain organs, for example rat brain, to induction of cancer by N-nitroso-N-alkyl-ureas, has been related to a low ability to remove O6-alkylguanine (O6AG) from DNA. It is therefore reasonable to ask why mouse brain, in which there is also a slow disappearance of O6AG from DNA after treatment with nitroso-alkyl-ureas, is not susceptible and why, in mice, thymus and lung are the main target organs. The explanation of the species difference could lie in the fact that replication of alkylated DNA is an essential event in initiation. If nitroso-alkyl-ureas had a greater inhibitory effect in some organs than in others, replication might be inhibited until after the O6AG had been removed, so preventing replication of DNA while still alkylated. This concept was tested by comparing the effect of N-nitroso-N-methyl-urea (NMU) on incorporation of [3H]TdR into DNA of relevant organs in Wistar rats and C57BL mice, and by determining ability to remove O6AG from DNA by measuring the alkyl acceptor protein (AAP) concentrations in these organs. No evidence was obtained that the AAP content was lower or inhibition of replication was less extensive in the organ of the species more susceptible to carcinogenesis than in the same organ of the less susceptible species. 相似文献
19.
Contribution of heterotrophic bacterial production to the carbon budget of the river Seine (France) 总被引:4,自引:0,他引:4
Bacterial activity was measured in the river Seine by two methods, 3H-thymidine incorporation into DNA and 3H-leucine incorporation into proteins. Both incorporation rates are characterized by low values upstream of Paris, a large increase just downstream of the outfall of the Achères treatment plant effluents, and then decreasing values further downstream. The covariation of both activities is demonstrated by the constancy of the molar ratio (leucine to thymidine incorporation rate) in the range of 6 to 8 for all the samples, except in the perturbed area where it is higher (15 to 35). These high values of molar ratio are linked to the introduction into the river of large sized bacteria (1 µm) with higher incorporation rates per cell or biomass unit than the small autochthonous bacteria (< 1 µm). Growth rates of large bacteria were on average 3.7 times higher than those of small bacteria. Bacterial production was calculated with experimentally determined conversion factors (0.5 × 1018 cells per mole of thymidine incorporated and 900 gC per mole of leucine incorporated) and by taking into account the activity of both size classes of bacteria measured through fractionation experiments (post-incubation filtration). Production estimated in the perturbed area downstream of Ach6res was very high, up to 60 µgC liter–1h–1 in the summer. Carbon consumption by bacteria in the area perturbed by the Ach6res effluents was calculated assuming a growth yield of 0.2 and compared to the load of biodegradable organic matter discharged by the treatment plant. In summer, an additional supply of organic matter is required to account for the intense bacterial activity, suggesting the importance of phytoplankton production in the carbon budget.
Offprint requests to: Pierre Servais 相似文献
20.
Effat Badr Mourad 《The Journal of cell biology》1965,24(2):267-276
Sterile root cultures from Nicotiana tabacum were grown with H3-thymidine added to the medium for various intervals. Incorporation of the labeled nucleoside into nuclear DNA occurred in a fraction of the nuclei which increased with time. In addition, the cytoplasm of all cells incorporated enough tritium to be readily detected by autoradiography. The tritium was not removed by hydrolysis in 1 N HCl at 60°C for 10 minutes, but was removed by digestion in a DNase solution which also removed nuclear DNA. The amount of tritium in the cytoplasm increased during the first 2 hours, but did not appear to increase significantly during the following 5 hours. If the roots were transferred to unlabeled medium after 2 hours, the label was diluted faster than expected by growth without turnover of the labeled component. If FUdR was added to the unlabeled medium, the depletion occurred faster during the first 6 hours, but later appeared to level off so that at 10 hours these cultures did not differ from those incubated without FUdR. However, the addition of an excess of unlabeled carrier had no effect on the rate of depletion of the cytoplasmic label. Actinomycin D, which inhibited the incorporation of H3-cytidine into RNA in the root tips, had no effect on the incorporation of H3-thymidine into the cytoplasmic component. However, Mitomycin C or a high concentration of deoxyadenosine inhibited the incorporation of H3-thymidine into the cytoplasmic component as well as into the nuclear DNA. It is concluded that H3-thymidine is incorporated into a cytoplasmic fraction which has the characteristics of DNA, with a measurable rate of turnover. This fraction is synthesized regardless of whether or not the nucleus is synthesizing DNA. Although the function of cytoplasmic fraction is not yet known, it does not appear to be that of supplying precursors for the synthesis of the nuclear DNA. 相似文献