Structure and Function of Suppressor tRNAs in Higher Eukaryote

Hatfield, Dolph L.; Smith, David W.; Lee, Byeong J.; Worland, Peter J.; Oroszlan, Stephen

doi:10.3109/10409239009090606

Cited by 57 publications

(21 citation statements)

References 233 publications

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“…However, these and other amino acids likely constitute a minor fraction as they were not observed even after targeted mass-spectrometry analysis. The observed readthrough across all stop codon types was unexpected, but nonsense suppression across all three stop codons has been reported in mammalian systems (Beier and Grimm, 2001; Hatfield et al, 1990). Recognition of all three stop codons by eRF1 is also consistent with our finding of promiscuous readthrough (Kisselev et al, 2003).…”

Section: Discussionmentioning

confidence: 67%

Programmed Translational Readthrough Generates Antiangiogenic VEGF-Ax

Eswarappa

Potdar

Koch

et al. 2014

Cell

189

288

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SUMMARY Translational readthrough, observed primarily in less complex organisms from viruses to Drosophila, expands the proteome by translating select transcripts beyond the canonical stop codon. Here we show that vascular endothelial growth factor-A (VEGFA) mRNA in mammalian endothelial cells undergoes programmed translational readthrough (PTR) generating VEGF-Ax, an isoform containing a unique 22-amino acid C-terminus extension. A cis-acting element in the VEGFA 3′UTR serves a dual function, not only encoding the appended peptide, but also directing the PTR by decoding the UGA stop codon as serine. Heterogeneous nuclear ribonucleoprotein (hnRNP) A2/B1 binds this element and promotes readthrough. Remarkably, VEGF-Ax exhibits anti-angiogenic activity in contrast to the pro-angiogenic activity of VEGF-A. Pathophysiological significance of VEGF-Ax is indicated by robust expression in multiple human tissues, but depletion in colon adenocarcinoma. Furthermore, genome-wide analysis revealed AGO1 and MTCH2 as authentic readthrough targets. Overall, our studies reveal a novel protein-regulated PTR event in a vertebrate system.

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Section: Discussionmentioning

confidence: 67%

Programmed Translational Readthrough Generates Antiangiogenic VEGF-Ax

Eswarappa

Potdar

Koch

et al. 2014

Cell

189

288

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“…It was in fact the sequence homology of this region to other proteins known to undergo readthrough that led to the initial suggestion of readthrough by Miller et aL (1988a). Although it is attractive to speculate that this difference in stop codon context may be a major factor in the suppression of the aphid-transmissibility phenotype, there is no compelling evidence that the stop codon context is critical in eukaryotic readthrough (Hatfield et al, 1990). It is also interesting that we have found through sequence analysis that the Cp-29K 'intergenic' region (including the entire proline-rich region) is identical in PEMV aphid nontransmissible strains and in strains propagated solely by aphid transmission.…”

Section: Analysis Of Pemv Structural Proteinsmentioning

confidence: 97%

The Nucleotide Sequence and Luteovirus-like Nature of RNA 1 of an Aphid Non-transmissible Strain of Pea Enation Mosaic Virus

Demler

Zoeten

1991

Journal of General Virology

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An examination of the genomic strategy of pea enation mosaic virus (PEMV) RNA 1 has verified strong organizational and sequence relationships between PEMV and the beet western yellows-potato leafroll luteovirus subgroup. Sequence analysis of RNA 1 demonstrated five predominant open reading frames (ORFs). The extreme 5' ORF encodes a 34K product of unknown function. The second ORF encodes an 84K product which overlaps 90% of ORF l (in a unique reading frame) and is expressed by internal initiation beginning at the second start codon from the 5' terminus. This protein contains a protease-like motif characteristic of serine-and cysteine-based proteases, suggesting involvement in post-translational processing of viral translation products. The third ORF is characterized by a number of RNA polymerase motifs and a helicase-like motif typical of RNA-dependent RNA polymerases. It overlaps (out of frame) the ORF 2 product and is proposed to be expressed by a frameshift fusion of the ORF 2 and ORF 3 products. The fourth ORF encodes the viral coat protein, and is immediately followed in frame by a 33K ORF thought to represent the aphid transmission subunit of the PEMV virion. Northern blot analysis of polysomeassociated RNA suggests that both products are expressed from an 1800 nucleotide subgenomic mRNA, with the 33K product expressed as a readthrough fusion with the coat protein monomer.

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“…They can insert an amino acid in response to a term ination codon. A num ber of suppresso r tRNAs have been identified in prokaryot es and eukaryotes such as yeast and Caenorha bditis elegans (Hatfield et al, 1990). Restorati on of the norm al phenotype by suppresso r tRNAs will depend on the nature of the amino acid substituted at the position of the nonsense codon and the efficiency of the translatio nal bypass.…”

Section: Introduction X Er O D Er M a Pigm En T O S U M (Xp) Is An Aumentioning

confidence: 99%

Partial Functional Correction of Xeroderma Pigmentosum Group A Cells by Suppressor tRNA

Panchal¹,

Wang²,

McDermott³

et al. 1999

Human Gene Therapy

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Genetic diseases are often caused by nonsense mutations. The resulting defect in protein translation can be restored by expressing suppressor tRNA in the mutant cells. Our goal was to demonstrate both protein restoration and phenotypic correction using these small transgenes. Functional activity of an arginine opal suppressor tRNA in cells expressing a nonsense mutated GFP gene was demonstrated by restored fluorescence. This suppressor tRNA was expressed in xeroderma pigmentosum group A cells, containing a homozygous nonsense mutation at Arg-207 in the XPA complementing gene. The transfected XPA cell population showed a twofold increase in cell survival after UV irradiation as determined by colony-forming assays compared with cell populations without the suppressor tRNA gene. The UV doses required for 37% survival of XP cells and XP cells expressing the suppressor tRNA were 0.6 and 1.2 J/m2. A similar twofold increase in the reactivation of UV-irradiated plasmid DNA was observed in XP cells expressing the suppressor tRNA. However, there was no detectable increase in XPA protein levels. Several potential limitations of this approach exist, including the availability of mutant RNA transcripts, the efficiency of suppression by the suppressor tRNA, and the abundance and availability and continued expression of the suppressor tRNA. The unique feature of this study is the relatively small size (88 bp) of the suppressor tRNA. Small-sized suppressor tRNAs can be synthetically constructed and subcloned into different viral vectors for delivery into the target cells. This approach may be useful for other genetic diseases caused by nonsense mutations.

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Structure and Function of Suppressor tRNAs in Higher Eukaryote

Cited by 57 publications

References 233 publications

Programmed Translational Readthrough Generates Antiangiogenic VEGF-Ax

Programmed Translational Readthrough Generates Antiangiogenic VEGF-Ax

The Nucleotide Sequence and Luteovirus-like Nature of RNA 1 of an Aphid Non-transmissible Strain of Pea Enation Mosaic Virus

Partial Functional Correction of Xeroderma Pigmentosum Group A Cells by Suppressor tRNA

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