The effects of actinomycin D and puromycin on morphogenesis in amphibian eggs and Acetabularia mediterranea

Brachet, Jean; Denis, Hélène; Vitry, F. De

doi:10.1016/0012-1606(64)90033-8

Cited by 168 publications

(26 citation statements)

References 44 publications

Supporting

Mentioning

Contrasting

Unclassified

Order By: Relevance

“…In the presence of actinomycin D embryos develop up to the MBT, whereas this development is blocked by puromycin or cycloheximide (3,4). Similar results have been obtained for oocyte maturation, which is independent of new transcription but requires de novo translation (5).…”

supporting

confidence: 72%

Cloning by differential screening of a Xenopus cDNA coding for a protein highly homologous to cdc2.

Paris

Guellec

Couturier

et al. 1991

Proc. Natl. Acad. Sci. U.S.A.

191

View full text Add to dashboard Cite

Fertilization ofXenopus laevis eggs triggers a period of rapid cell division comprising 12 nearly synchronous mitoses. Protein synthesis is required for these divisions, and new proteins appear after fertilization. Others proteins, however, which are synthesized in the unfertilized egg, are no longer made in the early embryo. To identify such proteins, a differential screen of an egg cDNA library gave nine clones corresponding to mRNAs that are deadenylylated soon after fertilization. The sequence ofone of these clones (Egi) revealed a high homology to p34Wdc2 the kinase subunit of maturationpromoting factor. Only 12 amino acids in the deduced amino acid sequence were unique to Egl when its sequence was compared to all other known examples of cdc2. Despite this strong similarity, however, Egl was unable to complement a yeast cdc2-mutant in Schizosaccharomyces pombe or a cdc28 mutant of Saccharomyces cerevisiae. Four Egl transcripts, two major and two minor, were found in Xenopus oocytes and early embryos. These RNAs appeared very early (stage I) in oogenesis and their level remained constant until the midlastfula transition, at which time they dedined. Egi RNA is found in the poly(A)+ fraction of oocytes only between the time of meiotic maturation and fertilization-that is to say, in the unfertilized egg. At fertilization the RNA loses its poly(A) tail and at the same time leaves the polyribosomes.

show abstract

supporting

confidence: 72%

Cloning by differential screening of a Xenopus cDNA coding for a protein highly homologous to cdc2.

Paris

Guellec

Couturier

et al. 1991

Proc. Natl. Acad. Sci. U.S.A.

191

View full text Add to dashboard Cite

show abstract

“…The onset of transcription is clearly detected only after cleavage 12 at a stage called the midblastula transition (23). In the presence of dactinomycin, embryos develop up to the midblastula transition, whereas this development is blocked by puromycin or cycloheximide (3,20). Similar results have been obtained for maturation, which is independent of new transcription but requires de novo translation (15).…”

supporting

confidence: 71%

Cloning by differential screening of a Xenopus cDNA that encodes a kinesin-related protein.

Guellec¹,

Paris²,

Couturier³

et al. 1991

Mol. Cell. Biol.

134

View full text Add to dashboard Cite

By differential screening of a Xenopus egg cDNA library, we selected nine clones (Egl to Eg9) corresponding to mRNAs which are deadenylated and released from polysomes soon after fertilization. The sequence of one of these clones (Eg5) revealed that the corresponding protein has the characteristic features of a kinesin-related protein. More specifically, Eg5 was found to be nearly 30% identical to a kinesin-related protein encoded by bimc, a gene involved in nuclear division in Aspergillus nidulans.

show abstract

“…This might also partially explain the selective action of actinomycin on histone synthesis, the similarity of the biological effects of this antibiotic and histones [35,36], and competition between the two.…”

Section: Discussionmentioning

confidence: 99%

Differential Inhibition of Protein Synthesis in the Nucleus and Cytoplasm of HeLa Cells in the Presence of Actinomycin, Puromycin and Hydroxyurea

Zampetti‐Bosseler

Malpoix

Fievez

1969

European Journal of Biochemistry

View full text Add to dashboard Cite

A comparison has been made between the synthesis of histones, other nuclear proteins and cytoplasmic proteins after treatment of HeLa cells with substances known to alter their metabolic state : actinomycin, puromycin and hydroxyurea. The use of sensitive methods involving amino acid analysis and measurement of specific radioactivity has revealed differential responses to the action of the different inhibitors. Analysis of these findings as a function of the effects of these inhibitors on other metabolic pathways, suggests that control mechanisms or basic mode of synthesis of some of the histones may differ from that of the other proteins, and that DNA and histone synthesis are probably coordinated in some way yet to be elucidated.Since the hypothesis of genetic repression by histones was advanced [l], many authors have sought to characterize their synthesis [2]. The present investigation has been carried out in an attempt to determine whether histones are synthesized in exactly the same way as the other proteins of the cell or whether some biosynthetic parameter (either a feature of control or a basic mechanism) distinguishes them. Their site of synthesis may be nuclear [3--51, nucleolar [6, 71, or cytoplasmic [8]; experimental evidence a t present available does not indicate clearly which of these possibilities is correct. Nor is it yet known whether the synthesis of all the histones is concomitant with that of DNA [lo-151 or whether some histones are synthesized outside the "S" period[16]. Differential rates of turnover, during different stages of the cell cycle, have been observed for different histone fractions [17]. The way in which such biosynthetic mechanisms are controlled and coordinated remains to be discovered. I n studying the effects of actinomycin D, hydroxyurea and puromycin on the synthesis of histones, residual nuclear proteins and cytoplasmic proteins, we have hoped to throw some light upon these problems. I n some experiments, in order to detect differential mechanisms, F1 (lysine-rich) histones were distinguished from other histones. Parallel experiments also established the effects of similar concentrations of the inhibitors on DNA synthesis. MATERIALS AND METHODSHeLa cells were grown a t 37" as monolayers in Puromycin, at concentrations ranging from 1.5 to 25 pg/ml, or hydroxyurea (76pg/ml) were administered a t the same time as the radioisotope, for a period of I to 1.5 hours. I n the case of actinomycin (1 pg/ml), the antibiotic was administered for an initial "pretreatment" period of 3 hours prior to the addition to the medium of labeled amino acids or thymidine. When protein synthesis was to be analysed, HeLa cells were exposed to ~-[4,5-~H]lysine 5 pC/ml (specific activity 510 mC/mmole) or to L3H]pro1ine 5 pC/ml (specific activity 324 mC/mmole). For studies of DNA synthesis, [6-3H]thymidine 2 pC/ml (specific activity 3.0 C/mmole) was added. Radioisotopes were all obtained from the Radiochemical Centre (Amersham, Buckinghamshire, Great Britain). Preparation of Protein FractionsThe inc...

show abstract

The effects of actinomycin D and puromycin on morphogenesis in amphibian eggs and Acetabularia mediterranea

Cited by 168 publications

References 44 publications

Cloning by differential screening of a Xenopus cDNA coding for a protein highly homologous to cdc2.

Cloning by differential screening of a Xenopus cDNA coding for a protein highly homologous to cdc2.

Cloning by differential screening of a Xenopus cDNA that encodes a kinesin-related protein.

Differential Inhibition of Protein Synthesis in the Nucleus and Cytoplasm of HeLa Cells in the Presence of Actinomycin, Puromycin and Hydroxyurea

Contact Info

Product

Resources

About