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Structural basis for cooperative RNA binding and export complex assembly by HIV Rev

Nature Structural & Molecular Biology volume 17, pages 1337–1342 (2010)Cite this article

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Abstract

HIV replication requires nuclear export of unspliced viral RNAs to translate structural proteins and package genomic RNA. Export is mediated by cooperative binding of the Rev protein to the Rev response element (RRE) RNA, to form a highly specific oligomeric ribonucleoprotein (RNP) that binds to the Crm1 host export factor. To understand how protein oligomerization generates cooperativity and specificity for RRE binding, we solved the crystal structure of a Rev dimer at 2.5-Å resolution. The dimer arrangement organizes arginine-rich helices at the ends of a V-shaped assembly to bind adjacent RNA sites and structurally couple dimerization and RNA recognition. A second protein-protein interface arranges higher-order Rev oligomers to act as an adaptor to the host export machinery, with viral RNA bound to one face and Crm1 to another, the oligomers thereby using small, interconnected modules to physically arrange the RNP for efficient export.

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Figure 1: Rev domain structure and protein dimerization.
Figure 2: Overall structure of the Rev dimer and monomer.
Figure 3: The Rev dimer interface mediates cooperative RNA recognition.
Figure 4: Arrangement of the Rev oligomer and model of the Rev–RRE RNP and Crm1 interaction.
Figure 5: Model for oligomerization-mediated cooperative assembly.

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Acknowledgements

We are grateful for assistance with crystallography and structure determination from Z. Newby, P. Egea, J. Finer-Moore, R. Stroud, M. Pufall, J.J. Miranda, D.Y. Kim, B. Kaiser, M. Brewer, L. Beamer, J. Holton and G. Meigs. We also thank D. Booth, B. Jayaraman, John Gross, Y. Cheng, R. Andino, G. Narlikar and H. Malik for helpful discussions and critical reading of the manuscript. M.D.D. was supported by a Howard Hughes Medical Institute predoctoral fellowship.

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Authors and Affiliations

  1. Chemistry and Chemical Biology Graduate Program, University of California, San Francisco, San Francisco, California, USA

    Matthew D Daugherty

  2. Department of Biochemistry and Biophysics, University of California, San Francisco, San Francisco, California, USA.,

    Bella Liu & Alan D Frankel

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  1. Matthew D Daugherty
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Contributions

M.D.D. and A.D.F. designed experiments and analyzed data. M.D.D. conducted RNA-binding and SEC experiments. M.D.D. and B.L. purified and crystallized Rev and collected diffraction data. M.D.D. solved and refined the structure. M.D.D. and A.D.F. wrote the manuscript.

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Correspondence to Alan D Frankel.

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The authors declare no competing financial interests.

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Supplementary Figures 1–4 and Supplementary Methods (PDF 1585 kb)

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Daugherty, M., Liu, B. & Frankel, A. Structural basis for cooperative RNA binding and export complex assembly by HIV Rev. Nat Struct Mol Biol 17, 1337–1342 (2010). https://doi.org/10.1038/nsmb.1902

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