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G-quadruplexes regulate Epstein-Barr virus–encoded nuclear antigen 1 mRNA translation

Nature Chemical Biology volume 10, pages 358–364 (2014)Cite this article

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Abstract

Viruses that establish latent infections have evolved unique mechanisms to avoid host immune recognition. Maintenance proteins of these viruses regulate their synthesis to levels sufficient for maintaining persistent infection but below threshold levels for host immune detection. The mechanisms governing this finely tuned regulation of viral latency are unknown. Here we show that mRNAs encoding gammaherpesviral maintenance proteins contain within their open reading frames clusters of unusual structural elements, G-quadruplexes, which are responsible for the cis-acting regulation of viral mRNA translation. By studying the Epstein-Barr virus–encoded nuclear antigen 1 (EBNA1) mRNA, we demonstrate that destabilization of G-quadruplexes using antisense oligonucleotides increases EBNA1 mRNA translation. In contrast, pretreatment with a G-quadruplex–stabilizing small molecule, pyridostatin, decreases EBNA1 synthesis, highlighting the importance of G-quadruplexes within virally encoded transcripts as unique regulatory signals for translational control and immune evasion. Furthermore, these findings suggest alternative therapeutic strategies focused on targeting RNA structure within viral ORFs.

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Figure 1: Identification and characterization of G-quadruplex structure within the EBNA1 GAr mRNA.
Figure 2: Codon modification enhances EBNA1 mRNA translation by destabilizing G-quadruplex structures.
Figure 3: The antisense oligonucleotide AS2 destabilizes g4-EBNA1 G-quadruplexes to enhance EBNA1 expression.
Figure 4: The small molecule PDS stabilizes g4-EBNA1 G-quadruplex structure and inhibits EBNA1 translation.
Figure 5: A schematic summarizing the translational control of EBNA1 mRNA by cis-regulatory elements, G-quadruplexes, leading to immune evasion.

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Change history

  • 04 April 2014

    In the version of this article initially published online, the third subheading in the Results subsection was incorrectly written as "Destabilizing G-quadruplexes mRNA translation"; it should read "Destabilizing G-quadruplexes enhances mRNA translation". Also, 'Cambridge Research Institute', as listed under the Affiliations, should be 'Cambridge Institute', and the last line of the Acknowledgments, originally written as "P.M. is supported by Cancer Research UK.", should read "The Balasubramanian group is supported by a program grant funded by Cancer Research UK." These errors have been corrected for the print, PDF and HTML versions of this article.

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Acknowledgements

We thank J. Shine and R. Tellam for valuable discussions, D. Hoang-Le and G. Wei for technical assistance and M. Di Antonio for providing PDS. Project grants (496684 and APP1005091) from the National Health and Medical Research Council (NHMRC) of Australia supported this research. J.T. was supported with a NHMRC Career Development Award Fellowship (no. 496712). R.K. is supported with a NHMRC Senior Principal Research Fellowship (APP1002476). The Balasubramanian group is supported by a program grant funded by Cancer Research UK.

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Authors and Affiliations

  1. Department of Chemistry, University of Cambridge, Cambridge, UK

    Pierre Murat & Shankar Balasubramanian

  2. Department of Immunology, Tumour Immunology, Clive Berghofer Cancer Research Centre, QIMR Berghofer Medical Research Institute, Brisbane, Queensland, Australia

    Jie Zhong, Lea Lekieffre, Rajiv Khanna & Judy Tellam

  3. QIMR Centre for Immunotherapy and Vaccine Development, QIMR Berghofer Medical Research Institute, Brisbane, Queensland, Australia

    Jie Zhong, Lea Lekieffre, Rajiv Khanna & Judy Tellam

  4. Centre for Synchrotron Science, Monash University, Melbourne, Victoria, Australia

    Nathan P Cowieson

  5. Genome Biology Department, The John Curtin School of Medical Research, The Australian National University, Canberra, Australian Capital Territory, Australia

    Jennifer L Clancy & Thomas Preiss

  6. Cambridge Institute, Cancer Research UK, Li Ka Shing Center, Cambridge, UK

    Shankar Balasubramanian

  7. School of Clinical Medicine, The University of Cambridge, Addenbrooke's Hospital, Hills Road, Cambridge, UK

    Shankar Balasubramanian

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Contributions

P.M. designed and undertook the biophysical experiments and computational analyses and analyzed the data; J.T. and L.L. designed and generated the constructs; J.Z. performed the antigen presentation studies; N.P.C. and J.T. performed the SAXS experiments; J.L.C., J.T. and T.P. performed and analyzed the polysome experiments; S.B. and R.K. provided technical and scientific advice and comments on the manuscript. J.T. conceived the project, undertook the biochemical and immunological experiments and analyzed the data. J.T. and P.M. wrote the manuscript.

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Correspondence to Judy Tellam.

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Supplementary Results, Supplementary Figures 1–18, Supplementary Tables 1 and 2 and Supplementary Note. (PDF 26519 kb)

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Murat, P., Zhong, J., Lekieffre, L. et al. G-quadruplexes regulate Epstein-Barr virus–encoded nuclear antigen 1 mRNA translation. Nat Chem Biol 10, 358–364 (2014). https://doi.org/10.1038/nchembio.1479

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