1,721,049 research outputs found
The 5' untranslated region of mRNA for ribosomal protein S19 is involved in its translational regulation during Xenopus development
No full textDuring Xenopus development, the synthesis of ribosomal proteins is regulated at the translational level. To identify the region of the ribosomal protein mRNAs responsible for their typical translationa- behavior, we constructed a fused gene in which the upstream sequences (promoter) and the 5' untranslated sequence (first exon) of the gene coding for Xenopus ribosomal protein S19 were joined to the coding portion of the procaryotic chloramphenicol acetyltransferase (CAT) gene deleted of its own 5' untranslated region. This fused gene was introduced in vivo by microinjection into Xenopus fertilized eggs, and its activity was monitored during embryogenesis. By analyzing the pattern of appearance of CAT activity and the distribution of the S19-CAT mRNA between polysomes and messenger ribonucleoproteins, it was concluded that the 35-nucleotide-long 5' untranslated region of the S19 mRNA is able to confer to the fused S19-CAT mRNA the translational behavior typical of ribosomal proteins during Xenopus embryo development
The Xenopus intron-encoded U17 snoRNA is produced by exonucleolytic processing of its precursor in oocytes
No full textU17 is a small nucleolar RNA encoded in the introns of the Xenopus laevis gene for ribosomal protein S7 (formerly S8, see Note). To study the mechanisms involved in its in vivo processing from S7 transcripts, various in vitro synthesized RNAs embedding a U17 sequence have been microinjected into the germinal vesicle of Xenopus oocytes and their processing analysed. In particular, the Xenopus U17 gene copies a and f and a U17 gene copy from the pufferfish Fugu rubripes have been used. Information about the nature of the processing activities involved in U17 RNA maturation have been sought by injecting transcripts protected from exonucleolytic attack at their 5'-end by capping and/or lengthened at their 3'-end by polyadenylation. The results obtained indicate that U17 RNA processing is a splicing-independent event and that it is mostly or entirely due to exonucleolytic degradation at both the 5'- and 3'-ends of the precursor molecules. Moreover, it is concluded that the enzymes involved are of the processive type. It is suggested that the apparatus for U17 RNA processing is that responsible for the degradation of all excised and debranched introns. Protection from exonucleolytic attack, due to the tight structure and/or to the binding of specific proteins, would be the mechanism by which U17 RNA is produced
Coordinate translational regulation in the syntheses of elongation factor 1 alpha and ribosomal proteins in Xenopus laevis
No full textThe regulation of the synthesis of elongation factor 1alpha (EF-1alpha) in Xenopus laevis has been analyzed from the point of view of translational control. The 5' end of EF-1alpha mRNA, examined by primer extension, revealed the presence of a terminal pyrimidine tract that is characteristic of ribosomal protein mRNAs (rp-mRNAs). We have then compared the translation pattern of EF-1alpha and rp-mRNAs during Xenopus embryogenesis and in Xenopus cultured cells during growth rate changes. In Xenopus embryos EF-1alpha transcripts, that appear after midblastula transition, are initially mostly localized on mRNP and translationally inactive. Only later in embryogenesis, together with rp-mRNAs, they are gradually recruited on polysomes. Also in Xenopus cells B 3.2, EF-1alpha mRNA shows a distribution change similar to an rp-mRNA: part of it moves from polysomes to mRNP during serum deprivation and goes back on polysomes after restitution of serum to the culture. Moreover EF-1alpha mRNA, similarly to rp-mRNAs, is always localized on mRNP or fully loaded on polysomes but never on small polysomes. Therefore EF-1alpha mRNA for structural features and translation behavior can be included in the 'regulatory' group of rp-mRNAs
Translational regulation of ribosomal protein synthesis in Xenopus cultured cells: mRNA relocation between polysomes and RNP during nutritional shifts
No full textTranslational control of ribosomal protein mRNA was analyzed in a Xenopus cell line during growth-rate changes induced by serum deprivation and readdition. After being transferred into serum-free medium, the cells rapidly decrease their DNA, RNA and protein synthesis, while addition of serum to the culture after a few hours of deprivation causes a rapid recovery. During these growth-rate changes, we observed a shift in ribosomal protein mRNA distribution between polysomes and RNP. The proportion of mRNA on polysomes for the four ribosomal proteins analyzed changed from 70-80% during rapid growth to 25-35% during the downshift and back to 70-80% after the upshift. Northern blot analysis showed that ribosomal protein mRNA level was constant during the shifts even in the presence of the transcriptional inhibitor actinomycin D. This indicates that the distribution changes were due to a reversible transfer of ribosomal protein mRNA between polysomes and RNP without altering mRNA stability. We have also compared the kinetics of ribosomal protein mRNA distribution changes with the kinetics of the changes in the partition of ribosomes between free monomers and polysomes. The results obtained show that the change in ribosomal protein mRNA localization is very fast, allowing short-term adjustments of ribosome synthesis rate. Moreover, our observations are consistent with the hypothesis that the amount of free ribosomes present in the cell could affect ribosomal protein mRNA utilization
Aspects of regulation of ribosomal protein synthesis in Xenopus laevis - Review
No full textThe work carried out in the authors' laboratories on the structure and expression of ribosomal protein genes in Xenopus is reviewed, with some comparisons with other systems. These genes form a class that shares several structural features, especially in the region surrounding the 5′ ends. These similar structures appear to be involved in coregulated expression that is attained at various regulatory levels: transcriptional, transcript processing and stability, and translational. Particular attention is paid here to the one operating at the translational level, which has been studied during Xenopus oogenesis and embryogenesis, and also during nutritional changes of Xenopus cultured cells. This regulation, which responds to the cellular need for new ribosomes, operates by changing the fraction of rp-mRNA engaged on polysomes, leaving each translated rp-mRNA molecule always fully loaded with ribosomes. Responsible for this translational behaviour is the typical 5′UTR, which characterizes all rp-mRNAs analyzed up to now, and that can bind in vitro some proteins, putative trans-acting factors for this translational regulation. © 1994 Kluwer Academic Publishers
Human ribosomal protein L4: Cloning and sequencing of the cDNA and primary structure of the protein
No full textThe cloning and sequencing of a cDNA for human ribosomal protein L4 is reported. The corresponding mRNA has a very short 5' untranslated region initiating with a sequence of 12 pyrimidines, characteristic of all vertebrate ribosomal protein mRNAs. The deduced amino acid sequence shows that human ribosomal protein L4 has 425 amino acid residues and a calculated molecular mass of 47821 Da. Comparison with the homologous counterparts of Xenopus, Drosophila and yeast shows that this protein has a very conserved amino-terminus region and an extremely divergent carboxyl-terminus portion
Analysis of mRNAs under translational control during Xenopus embryogenesis: Isolation of new ribosomal protein clones
No full textWe have analyzed several randomly selected mRNAs, of the relatively abundant category, on the basis of maternal or zygotic origin and translational efficiency at different developmental stages. For this purpose, clones from a Xenopus embryo cDNA library were hybridized with cDNA probes prepared with poly(A) + RNA from polysomes and from mRNPs of embryos at different stages. The results obtained indicate that the majority of the relatively abundant MRNAs (38 out of 61) is subject to some kind of translational regulation during embryogenesis. Moreover, 30 clones have been selected as corresponding to mRNAs that behave, from the point of view of transcriptional and translational regulation, similarly to previously studied ribosomal protein (r-protein) mRNAs. Sequence analysis of 20 of these selected cDNAs has shown that half of them are in fact homologous to already sequenced r-protein mRNAs. Unexpectedly we have found that also the mRNA for alpha-cardiac actin and another mRNA homologous to creatine kinase M mRNA have a similar translational regulation during embryogenesis
Translational control of terminal oligopyrimidine mRNAs requires a specific regulator
No full textTerminal oligopyrimidine (TOP) mRNAs are a group of messengers translationally regulated according to the growth status of the cell. Two hypotheses have been proposed for the mechanism of the regulation: (i) there is a specific translational regulator which can reversibly alter TOP-mRNA structure, (ii) a component of the general translational apparatus can specifically affect the translation of TOP-mRNAs. To verify one of the two hypotheses we induced a partial inhibition of translation initiation in Xenopus cultured cells and analyzed the effect on TOP-mRNA translation. Our results suggest that a specific regulator is necessary to explain the translational control of these of mRNAs
XENOPUS-LAEVIS RIBOSOMAL-PROTEIN S11 - CLONING AND SEQUENCING OF THE CDNA AND PRIMARY STRUCTURE OF THE PROTEIN
No full textINDIVIDUAL VARIABILITY IN THE TRANSLATIONAL REGULATION OF RIBOSOMAL-PROTEIN SYNTHESIS IN XENOPUS-LAEVIS
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