In Xenopus laevis, the product of a developmentally regulated mRNA is structurally and functionally homologous to a Saccharomyces cerevisiae protein involved in translation fidelity.

Tassan, Jean‐Pierre; Guellec, Katherine Le; Kress, Michel; Faure, Michel; Camonis, Jacques; Jacquet, Michel

doi:10.1128/mcb.13.5.2815

Cited by 34 publications

(24 citation statements)

References 40 publications

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“…One possible way of testing this model is to examine whether increasing the levels of the translation release factors leads to suppression of selenocysteine incorporation. To perform this experiment, we co-transfected individual reporter constructs together with expression constructs bearing the genes encoding eRF1 and eRF3 (29,30). Comparison with the control construct pBLUGA revealed that there was little specific repression of selenocysteine incorporation associated with the overproduction of the eRFs (Fig.…”

Section: Resultsmentioning

confidence: 99%

Eukaryotic Selenocysteine Incorporation Follows a Nonprocessive Mechanism That Competes with Translational Termination

Nasim¹,

Jaenecke²,

Beldüz³

et al. 2000

Journal of Biological Chemistry

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The synthesis of eukaryotic selenoproteins involves the recoding of an internal UGA codon as a site for selenocysteine incorporation. This recoding event is directed by a selenocysteine insertion sequence in the 3-untranslated region. Because UGA also functions as a signal for peptidyl-tRNA hydrolysis, we have investigated how the rates of translational termination and selenocysteine incorporation relate to cis-acting elements in the mRNA as well as to trans-acting factors in the cytoplasm. We used cis-elements from the phospholipid glutathione peroxidase gene as the basis for this work because of its relatively high efficiency of selenocysteine incorporation. The last two codons preceding the UGA were found to exert a far greater influence on selenocysteine incorporation than nucleotides downstream of it. The efficiency of selenocysteine incorporation was generally much less than 100% but could be partially enhanced by concomitant overexpression of the tRNA Sec gene. The combination of two or three UGA codons in one reading frame led to a dramatic reduction in the yield of full-length protein. It is therefore unlikely that multiple incorporations of selenocysteine are processive with respect to the mode of action of the ribosomal complex binding to the UGA site. These observations are discussed in terms of the mechanism of selenoprotein synthesis and its ability to compete with termination at UGA codons.

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Section: Resultsmentioning

confidence: 99%

Eukaryotic Selenocysteine Incorporation Follows a Nonprocessive Mechanism That Competes with Translational Termination

Nasim¹,

Jaenecke²,

Beldüz³

et al. 2000

Journal of Biological Chemistry

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“…Plasmid Cl2Al-454,480-559 was made by digesting plasmid Cl2Al-454 with NheI (just upstream of the CPE) and EcoRI (located downstream in the vector); the larger fragment, which contained only a few C12 sequences upstream of the CPE, was filled in with Klenow enzyme and relegated. Plasmids Cl1-3'UTR, Cl1A1-179, CliA1-219, and Cl1lA235-5OSAUUAAA were constructed by PCR using the following oligonucleotides and a subclone of the Cli cDNA (38) sequences of this region (Cl11-179, C11A287-468, and Clil1-219; Fig. 1).…”

Section: Methodsmentioning

confidence: 99%

Further analysis of cytoplasmic polyadenylation in Xenopus embryos and identification of embryonic cytoplasmic polyadenylation element-binding proteins.

Simon¹,

Richter²

1994

Mol. Cell. Biol.

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Early development in Xenopus laevis is programmed in part by maternally inherited mRNAs that are synthesized and stored in the growing oocyte. During oocyte maturation, several of these messages are translationally activated by poly(A) elongation, which in turn is regulated by two cis elements in the 3' untranslated region, the hexanucleotide AAUAAA and a cytoplasmic polyadenylation element (CPE) consisting of UUUUUAU or similar sequence. In the early embryo, a different set of maternal mRNAs is translationally activated. We have shown previously that one of these, C12, requires a CPE consisting of at least 12 uridine residues, in addition to the hexanucleotide, for its cytoplasmic polyadenylation and subsequent translation (R. Simon, J.-P. Tassan, and J. D. Richter, Genes Dev. 6:2580Dev. 6: -2591Dev. 6: , 1992 for cytoplasmic polyadenylation at times up to the gastrula stage. This RNA also required a poly(U) CPE for cytoplasmic polyadenylation in embryos, but in this case the CPE consisted of 18 uridine residues. In addition, the timing and extent of cytoplasmic poly(A) elongation during early embryogenesis were dependent upon the distance between the CPE and the hexanucleotide. Further, as was the case with C12 RNA, Cli RNA contains a large masking element that prevents premature cytoplasmic polyadenylation during oocyte maturation. To examine the factors that may be involved in the cytoplasmic polyadenylation of both C12 and Cli RNAs, we performed UV cross-linking experiments in egg extracts. Two proteins with sizes of -36 and -45 kDa interacted specifically with the CPEs of both RNAs, although they bound preferentially to the C12 CPE. The role that these proteins might play in cytoplasmic polyadenylation is discussed.

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“…The gel was subsequently electrotransferred onto Hybond-P PVDF membrane (Amersham Pharmacia Biotech), and the membrane blocked overnight at 4°C in 1× phosphate-buffered saline (PBS) containing 3% milk, 2% Normal Goat Serum, and 0.1% Tween-20. Blots were probed with anti-eRF1 serum (a generous gift from Michel Philippe (Université de Rennes I); Tassan et al 1993) in blocking solution at room temperature for 60 min. Blots were washed three times for 15 min in 30 mL of 1× PBS containing 0.1% Tween-20.…”

Section: Western Blotmentioning

confidence: 99%

Stop codon suppression via inhibition of eRF1 expression

Carnes

Jacobson²,

Leinwand³

et al. 2003

RNA

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In humans, recognition of a stop codon by protein release factor eRF1 leads to release of the nascent peptide from the ribosome. Although efficient eRF1 activity is usually desirable, numerous pathologies result from eRF1 recognition of premature stop mutations in essential genes. In these cases, decreased eRF1 activity could increase readthrough of the premature stop codon, thereby making full-length protein. To broaden the means available to beneficially decrease eRF1 activity, we have targeted eRF1 mRNA using siRNAs and antisense oligonucleotides. We show that both eRF1-targeted siRNA and antisense oligonucleotides decrease eRF1 mRNA and eRF1 protein concentrations, and increase UAG readthrough in cultured human cells.

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In Xenopus laevis, the product of a developmentally regulated mRNA is structurally and functionally homologous to a Saccharomyces cerevisiae protein involved in translation fidelity.

Cited by 34 publications

References 40 publications

Eukaryotic Selenocysteine Incorporation Follows a Nonprocessive Mechanism That Competes with Translational Termination

Eukaryotic Selenocysteine Incorporation Follows a Nonprocessive Mechanism That Competes with Translational Termination

Further analysis of cytoplasmic polyadenylation in Xenopus embryos and identification of embryonic cytoplasmic polyadenylation element-binding proteins.

Stop codon suppression via inhibition of eRF1 expression

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