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Effect of a natural mutation in the 5′ untranslated region on the translational control of p53 mRNA

Oncogene volume 32, pages 4148–4159 (2013)Cite this article

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Abstract

Tumor-suppressor protein p53, the ‘guardian of the genome’, is critical in maintaining cellular homeostasis and genomic stability. Earlier, we have reported the discovery of internal ribosome entry sites (IRESs) within the p53 mRNA that regulate the translation of the full length and its N-terminal-truncated isoform, ΔN-p53. Polypyrimidine tract-binding protein (PTB) is an IRES trans-acting factor that positively regulates the IRES activities of both p53 isoforms by relocating from nucleus to the cytoplasm during stress conditions. Here we have demonstrated the putative contact points of PTB on the p53 IRES RNA. Studies on mutations that occur naturally in the 5′ untranslated region (5′ UTR) in p53 mRNA were lacking. We have investigated a naturally occurring C-to-T single-nucleotide polymorphism (SNP) first reported in human melanoma tumors. This SNP is at position 119 in the 5′ UTR of p53 mRNA and we demonstrate that it has consequences on the translational control of p53. Introduction of this SNP has led to decrease in cap-independent translation from p53 5′ UTR in bicistronic reporter assay. Further, the effects of this SNP on cap-independent translation have been studied in the context of p53 cDNA as well. Interestingly, the 5′ UTR with this SNP has shown reduced binding to PTB that can be corroborated to its weaker IRES activity. Previously, it has been shown that G2–M checkpoint, DNA-damaging stress and oncogenic insult favor IRES-mediated translation. Under similar conditions, we demonstrate that this SNP interferes with the enhancement of the IRES activity of the 5′ UTR. Taken together, the results demonstrate for the first time that SNP in the 5′ UTR of the p53 mRNA might have a role in translational control of this critical tumor-suppressor gene.

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Acknowledgements

We are grateful to Professor Robin Fahraeus (INSERM, France) for sharing the pcDNA3-GFP-hp-p53-5′ UTR-cDNA construct with us. Professor Julian Downward (London Research Institute, UK) and Professor Annapoorni Rangarajan (Department of Molecular Reproduction and Developmental Genetics, IISc) are duly acknowledged for sharing the pBabe-H-RasV12 construct and anti-Ras antibody. We thank Professor K Somasundaram for sharing the WWP-Luciferase construct and Professor KN Balaji for providing anti-caspase-3 antibody. We also thank the FACS facility (Division of Biological Sciences, IISc) headed by Dr Omana Joy for the acquisition of the flow-cytometric data. We thank the present and past SD Lab members for critical discussion of the work and Ranjitha Tatineni for her help with the phylogenetic analysis. DK and SA are supported by pre-doctoral fellowships from CSIR, India. This work was partly supported by an Indo-French grant from IFCPAR to SD.

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  1. A Sharathchandra and A Ponnuswamy: These authors contributed equally to this work.

Authors and Affiliations

  1. Department of Microbiology and Cell Biology, Indian Institute of Science, Bangalore, India

    D Khan, A Sharathchandra, A Ponnuswamy, R Grover & S Das

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  1. D Khan
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Correspondence to S Das.

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Khan, D., Sharathchandra, A., Ponnuswamy, A. et al. Effect of a natural mutation in the 5′ untranslated region on the translational control of p53 mRNA. Oncogene 32, 4148–4159 (2013). https://doi.org/10.1038/onc.2012.422

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