Background Antizyme is a critical regulator of cellular polyamine levels due to its effect on polyamine transport and its ability to target ornithine decarboxylase for degradation. Antizyme expression is autoregulatory, through dependence on an unusual +1 translational frameshift mechanism that responds to polyamine levels. Results HEK293 cells were depleted of polyamines by treatment with an ornithine decarboxylase inhibitor, difluoromethylornithine (DFMO), and grown in the presence or absence of exogenous polyamines prior to the analysis of ribosomal frameshifting levels. Results obtained using an optimized dual luciferase assay system reveal a 10‐fold dynamic range of frameshifting, which correlates positively with polyamine addition. Polyamine addition to cells, which have not been pre‐treated with DFMO, also resulted in an increase in antizyme frameshifting but to a lesser degree (1.3 to 1.5‐fold). In addition, the constructs with the 3′ deletion were more responsive to stimulation by polyamine addition than those with the 5′ deletion. Conclusions The observed regulation of antizyme frameshifting demonstrates the efficiency of a polyamine homeostatic mechanism, and illustrates the utility of a quantifiable cell‐based assay for the analysis of polyamines or their analogues on translational frameshifting.

Ornithine decarboxylase antizyme

Translational frameshift

Polyamine

10.1046/j.1365-2443.2001.00477.x

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Citations (34)

AMD1 mRNA employs ribosome stalling as a mechanism for molecular memory formation

Nature (2018)

Martina M. Yordanova Gary Loughran Alexander V. Zhdanov Marco Mariotti Stephen J. Kiniry

Translational frameshift

Stop codon

Coding region

Shine-Dalgarno sequence

10.1038/nature25174

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Citations (76)

The nucleotide sequence of the first externally suppressible-1 frameshift mutant, and of some nearby leaky frameshift mutants.

The EMBO Journal (1983)

John F. Atkins Brian P. Nichols S. Thompson

Transversion

10.1002/j.1460-2075.1983.tb01590.x

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Citations (58)

Analysis of the roles of tRNA structure, ribosomal protein L9, and the bacteriophage T4 gene 60 bypassing signals during ribosome slippage on mRNA

Journal of Molecular Biology (2001)

Alan J. Herr Chad Nelson Norma M. Wills Raymond F. Gesteland John F. Atkins

Translational frameshift

Stop codon

P-site

T arm

Ribosomal binding site

Release factor

10.1006/jmbi.2001.4717

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Citations (81)

Identification of evolutionarily conserved non-AUG-initiated N-terminal extensions in human coding sequences

Nucleic Acids Research (2011)

Ivaylo P. Ivanov Andrew E. Firth Audrey Michel John F. Atkins Pavel V. Baranov

In eukaryotes, it is generally assumed that translation initiation occurs at the AUG codon closest to the messenger RNA 5' cap. However, in certain cases, initiation can occur at codons differing from AUG by a single nucleotide, especially the codons CUG, UUG, GUG, ACG, AUA and AUU. While non-AUG initiation has been experimentally verified for a handful of human genes, the full extent to which this phenomenon is utilized--both for increased coding capacity and potentially also for novel regulatory mechanisms--remains unclear. To address this issue, and hence to improve the quality of existing coding sequence annotations, we developed a methodology based on phylogenetic analysis of predicted 5' untranslated regions from orthologous genes. We use evolutionary signatures of protein-coding sequences as an indicator of translation initiation upstream of annotated coding sequences. Our search identified novel conserved potential non-AUG-initiated N-terminal extensions in 42 human genes including VANGL2, FGFR1, KCNN4, TRPV6, HDGF, CITED2, EIF4G3 and NTF3, and also affirmed the conservation of known non-AUG-initiated extensions in 17 other genes. In several instances, we have been able to obtain independent experimental evidence of the expression of non-AUG-initiated products from the previously published literature and ribosome profiling data.

Ribosome profiling

Coding region

Conserved sequence

Eukaryotic translation

Stop codon

10.1093/nar/gkr007

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Citations (212)

Electromyographic observations in the human larynx

Irish Journal of Medical Science (1971 -) (1968)

Francis J. Bonner John F. Atkins

10.1007/bf02958790

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Citations (4)

A three-way junction and constituent stem-loops as the stimulator for programmed −1 frameshifting in bacterial insertion sequence IS 911 1 1Edited by D. E. Draper

Journal of Molecular Biology (1999)

Charles C. Rettberg Marie Françoise Prère Raymond F. Gesteland John F. Atkins Olivier Fayet

Translational frameshift

Sequence (biology)

10.1006/jmbi.1999.2546

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Citations (45)

Nonsense Codons and Unassigned Codons

Encyclopedia of Molecular Biology (2002)

John F. Atkins K. Watanabe Shin‐ichi Yokobori

Nonsense

Encyclopedia

10.1002/047120918x.emb1014

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Citations (0)

A case for “StopGo”: Reprogramming translation to augment codon meaning of GGN by promoting unconventional termination (Stop) after addition of glycine and then allowing continued translation (Go)

RNA (2007)

John F. Atkins Norma M. Wills Gary Loughran Chih-yu Wu Krishna Parsawar

When a eukaryotic mRNA sequence specifying an amino acid motif known as 2A is directly followed by a proline codon, two nonoverlapping proteins are synthesized. From earlier work, the second protein is known to start with this proline codon and is not created by proteolysis. Here we identify the C-terminal amino acid of an upstream 2A-encoded product from Perina nuda picorna-like virus that is glycine specified by the last codon of the 2A-encoding sequence. This is an example of recoding where 2A promotes unconventional termination after decoding of the glycine codon and continued translation beginning with the 3′ adjacent proline codon.

Stop codon

Eukaryotic translation

10.1261/rna.487907

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Citations (118)