1,721,061 research outputs found
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
Complementarity of conserved sequence elements present in 28S ribosomal RNA and in ribosomal protein genes of Xenopus laevis and Xenopus tropicalis.
No full textThe sequence analysis of the L1 ribosomal protein (r-protein) gene of Xenopus laevis has revealed a strong homology in four out of the nine introns of the gene; this homology region spans 60 nucleotides (nt) with 80% homology [Loreni et al., EMBO J. 4 (1985) 3483-3488]. We have extended our analysis to X. tropicalis, a species which is closely related to X. laevis. Partial sequencing of the isolated L1 gene has revealed that these 60-nt homology regions are also present in at least two introns of the X. tropicalis L1 gene. Computer analysis has revealed that perfect nt sequence complementarity exists between 13 nt of this intron region and the 28S ribosomal RNA in a region which is conserved in all eukaryotes, suggesting a possible base-pairing interaction between these two sequences
TmRdb : Database per la famiglia dei TOP mRNA.
No full textRecent developments in the biochemistry and molecular biology studies, make obvious the crucial role of TOP mRNA in translational regulation mechanisms and their direct involvement in ribosomopathies and cancer. Currently are known only 93 TOP mRNA[1], we proposed to develop a web application that allows to identify new TOP mRNAs and their mapping into a relational database; this to help research development in this field and return a web based consultation tool in a user friendly and multidevice interface
EpCAMlow circulating tumor cells: Gold in the waste
Full text linkThe CellSearch® system which is still considered the gold standard for the enumeration of circulating tumor cells (CTC) utilizes antibodies against the epithelial cell adhesion molecule (EpCAM) for CTC enrichment. Recently, CTC discarded by the CellSearch® system due to their low EpCAM expression have been isolated and analyzed. We here sought to discuss technical and biological issues concerning the isolation and characterization of EpCAMlow CTC, highlighting the enormous potential of this subpopulation discarded by CellSearch®, which might instead reveal an unexpected clinical significance in tumor types where CTC enumeration has never been validated for prognostic and predictive purpose
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
The oncoprotein Myc controls the phosphorylation of S6 kinase and AKT through protein phosphatase 2A
No full textThis study focuses on the effects of Myc oncoprotein on the translational apparatus of the cell. Translation is an energy consuming process that involves a large number of accessory factors. The production of components of the protein synthesis machinery can be regulated at the transcriptional level by specific factors. It has been shown that the product of the oncogene Myc, a transcription factor frequently activated in cancer, can control translational activity through an increase in the transcription of the eIF4F complex components (eIF4E, eIF4AI, and eIF4GI). However, additional effects at the posttranslational level have also been described. For instance, it has been shown that Myc upregulation can induce mammalian target of rapamycin (mTOR)-dependent 4E-binding protein 1 (4E-BP1) hyperphosphorylation. We induced overexpression or inhibition of Myc through transfection of complementary DNA constructs or specific small interfering RNA in PC3 (prostate carcinoma) and HeLa (cervical carcinoma) cells. We have observed that overexpression of Myc causes an increase in 4E-BP1 phosphorylation and activation of protein synthesis. Unexpectedly, we detected a parallel decrease in the phosphorylation level of S6 kinase (in PC3 and HeLa) and AKT (in HeLa). We report evidence that these changes are mediated by an increase in protein phosphatase 2A activity
Translational regulation of terminal oligopyrimidine mRNAs induced by serum and amino acids involves distinct signaling events
No full textVarious mitogenic or growth inhibitory stimuli induce a rapid change in the association of terminal oligopyrimidine (TOP) mRNAs with polysomes. It is generally believed that such translational control hinges on the mammalian target of rapamycin (mTOR)-S6 kinase pathway. Amino acid availability affects the translation of TOP mRNAs, although the signaling pathway involved in this regulation is less well characterized. To investigate both serum- and amino acid-dependent control of TOP mRNA translation and the signaling pathways involved, HeLa cells were subjected to serum and/or amino acid deprivation and stimulation. Our results indicate the following. 1) Serum and amino acid deprivation had additive effects on TOP mRNA translation. 2) The serum content of the medium specifically affected TOP mRNA translation, whereas amino acid availability affected both TOP and non-TOP mRNAs. 3) Serum signaling to TOP mRNAs involved only a rapamycin-sensitive pathway, whereas amino acid signaling depended on both rapamycin-sensitive and rapamycin-insensitive but wortmannin-sensitive events. 4) Eukaryotic initiation factor-2 phosphorylation increased during amino acid deprivation, but not following serum deprivation. Interestingly, rapamycin treatment suggests a novel connection between the mTOR pathway and eukaryotic initiation factor-2 phosphorylation in mammalian cells, which may not, however, be involved in TOP mRNA translational regulation
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