Synthesis and transport of various RNA species in developing embryos of Xenopus laevis

Shiokawa, K.; Misumi, Yuko; Yasuda, Yoh; Nishio, Y.; Kurata, Shigeaki; Sameshima, M; Yamana, K.

doi:10.1016/0012-1606(79)90221-5

Cited by 39 publications

(15 citation statements)

References 34 publications

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“…1D). (18). Thus, it appears that in Xenopus embryos cordycepin inhibits the synthesis of all the types of RNAs.…”

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confidence: 99%

“…The soluble cytoplasmic fraction was mixed with 10 volumes of 0.1 M sodium acetate buffer that contained SDS and bentonite. RNA was extracted with chloroform-phenol at room temperature (17) and was precipitated from the final aqueous phase with 0.2 M NaCl and 3 volumes of ethanol, after which it was fractionated into poly(A)+RNA and poly(U)-Sepharose-void RNA on a column of poly(U)-Sepharose 4B (18). The lengths of the poly(A) sequences in the poly(A)+ RNA obtained may be longer than 20 nucleotides as reported in a previous paper (15).…”

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confidence: 99%

“…RNA was chromatographed on poly(A)-Sepharose 4B. The buffers used in the chromatograppy were the same as those used with poly(U)-Sepharose 4B (18). Poly(U)-Sepharose-void RNAs were fractionated by electrophoresis on 0.5 % agarose-2.2% polyacrylamide or 8 % polyacrylamide gels.…”

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confidence: 99%

“…Poly(A) + RNA and "oligo(U)+RNA" were fractionated on 3.8 % polyacrylamide gels containing 80 % formamide. The gels were sliced then treated overnight with 3N ammonia solution, after which radioactivity was measured in a scintillation spectrometer in Triton X100-toluene-DPO (18). cytoplasmic poly(A) + RNA more strongly than nuclear poly(A) + RNA because the smaller poly(A) + RNA appears to be mainly cytoplasmic (18).…”

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confidence: 99%

“…The gels were sliced then treated overnight with 3N ammonia solution, after which radioactivity was measured in a scintillation spectrometer in Triton X100-toluene-DPO (18). cytoplasmic poly(A) + RNA more strongly than nuclear poly(A) + RNA because the smaller poly(A) + RNA appears to be mainly cytoplasmic (18). The amount of labeled "oligo(U) + RNA" in the control cells was about one-fourth of the total labeled poly(A) + RNA.…”

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Effects of cordycepin on the synthesis of nuclear and cytoplasmic RNAs in embryonic cells of Xenopus laevis.

Shiokawa

Misumi

1983

Cell Struct. Funct.

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Cordycepin has been reported to interfere with mRNA polyadenylation, thereby inhibiting the post-transcriptional processing of mRNA (2,4,6,7,8,10). Cordycepin has been used as a specific inhibitor of mRNA polyadenylation in studies of mRNA metabolism in various developmental systems (3,9,11,16,21,22,23). In several adult-type cells, however, cordycepin has been claimed to inhibit the synthesis of both poly(A) + RNA and other RNA species (1,5,20).We examined the effects of cordycepin on cells from early stages of embryonic development. Early embryos of Xenopus laevis were dissociated and their isolated cells treated with cordycepin. Our results suggest that cordycepin inhibits the synthesis and cytoplasmic accumulation not only of mRNA but of other kinds of RNAs as well. MATERIALS AND METHODSIsolation and labeling of embryonic cells. Embryos of Xenopus laevis were obtained by the injection of a gonadotropic hormone. Embryos were dejellied and dissociated into cells at the gastrula (stage 10) (13) and late neurula (stage 20) stages then labeled for varying periods with [8-3H]guanosine (6 Ci/mmole), or with [3H(G)]3 '-deoxyadenosine (cordycepin) (9.5 Ci/mmole), in complete Stearns' medium as described by Shiokawa and Yamana (19).Cell fractionation. Labeled cells were homogenized in a Dounce-type homogenizer, 5 mM MgC12 and 50 mM Tris-HCl (pH 8.0). The homogenate was centrifuged for 5 min at 500 g. The resulting supernatant was used as the cytoplasmic fraction and the pellet as the nuclear fraction (18).Preparation of polysomes. Labeled cells were homogenized in a Dounce-type homoge-1 address:

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“…1D). (18). Thus, it appears that in Xenopus embryos cordycepin inhibits the synthesis of all the types of RNAs.…”

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confidence: 99%

mentioning

confidence: 99%

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confidence: 99%

mentioning

confidence: 99%

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confidence: 99%

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Effects of cordycepin on the synthesis of nuclear and cytoplasmic RNAs in embryonic cells of Xenopus laevis.

Shiokawa

Misumi

1983

Cell Struct. Funct.

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Small nuclear rnas in cellular growth and differentiation. I: Metabolic alterations seen in friend erythroleukemic cells

Kuncio

Goldstein

1981

Journal Cellular Physiology

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Electrophoretic analysis of near steady-state labeled nuclear RNA obtained from Friend virus-transformed murine erythroleukemic cells reveals the presence of at least 15 small nuclear RNAs (snRNAs) distinct from ribosomal 5.8S or 5S. Identical qualitative distributions were obtained from logarithmically growing, stationary-phase, and dimethyl sulfoxide-induced, terminally differentiated cultures, indicating the constitutive synthesis of all snRNAs regardless of the proliferative or differentiated state of the cells. However, several quantitative differences in nuclear snRNA levels were observed. Progression from rapidly growing to stationary-phase cultures was accompanied by the marked reduction in accumulation of all snRNAs except the 4.5S snRNAs. Particularly striking were the decreases in levels of U3 and the U1 group, snRNAs that are relatively abundant. Similar reductions were noted when cells were induced to differentiate, except that decreases in the levels of U2 and 4.5S were more dramatic than those seen for cells entering stationary-phase. The data thus demonstrate that snRNA levels may be regulated both in association with changes in proliferative capacity of cells and with changes in gene expression that occur during terminal differentiation.

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Determination of the rate of rRNA synthesis in Xenopus laevis triploid embryos produced by low‐temperature treatment

Tashiro¹,

Misumi²,

Shiokawa³

et al. 1983

J. Exp. Zool.

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Triploid embryos of Xenopus laevis were obtained by cold-temperature shocking of the fertilized eggs, and the rate of the ribosomal RNA (rRNA) synthesis was determined for comparison with that in diploid embryos. For this purpose, both triploid and diploid embryos were dissociated into cells at the neurula stage, and then labeled with (3H)uridine for varying lengths of time. The rate of rRNA synthesis, as estimated after determination of (3H)UTP specific radioactivity and the total label incorporation into the purified rRNA, was about 0.1 pg/cell/hr for both diploid and triploid embryo cells. Nuclei of triploid embryo cells contained three nucleoli of apparently similar sizes--an indication of the functioning of all the three rRNA gene clusters to a more or less similar extent. Also, rates of synthesis of 4S RNA and 5S RNA were determined: Both rates did not change appreciably between triploid and diploid embryo cells. Based on these results, it appears that transcription of these redundant genes occurs at a constant rate on a per cell basis irrespective of the presence of 1.5 times as many genes as the control.

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Synthesis and transport of various RNA species in developing embryos of Xenopus laevis

Cited by 39 publications

References 34 publications

Effects of cordycepin on the synthesis of nuclear and cytoplasmic RNAs in embryonic cells of Xenopus laevis.

Effects of cordycepin on the synthesis of nuclear and cytoplasmic RNAs in embryonic cells of Xenopus laevis.

Small nuclear rnas in cellular growth and differentiation. I: Metabolic alterations seen in friend erythroleukemic cells

Determination of the rate of rRNA synthesis in Xenopus laevis triploid embryos produced by low‐temperature treatment

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