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Direct ribosomal binding by a cellular inhibitor of translation

Nature Structural & Molecular Biology volume 13, pages 103–111 (2006)Cite this article

Abstract

During apoptosis and under conditions of cellular stress, several signaling pathways promote inhibition of cap-dependent translation while allowing continued translation of specific messenger RNAs encoding regulatory and stress-response proteins. We report here that the apoptotic regulator Reaper inhibits protein synthesis by binding directly to the 40S ribosomal subunit. This interaction does not affect either ribosomal association of initiation factors or formation of 43S or 48S complexes. Rather, it interferes with late initiation events upstream of 60S subunit joining, apparently modulating start-codon recognition during scanning. CrPV IRES–driven translation, involving direct ribosomal recruitment to the start site, is relatively insensitive to Reaper. Thus, Reaper is the first known cellular ribosomal binding factor with the potential to allow selective translation of mRNAs initiating at alternative start codons or from certain IRES elements. This function of Reaper may modulate gene expression programs to affect cell fate.

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Figure 1: Ribosomes restore Reaper's inhibitory activity.
Figure 2: Reaper binds ribosomes through direct interaction with the 40S subunit.
Figure 3: Reaper inhibits translation initiation and induces accumulation of 48S half-mers.
Figure 4: Reaper does not affect eIF2α phosphorylation or 43S complex formation.
Figure 5: Reaper-induced 48S half-mers inefficiently recognize the AUG start site.
Figure 6: Reaper induces differential pausing in bicistronic mRNA messages.
Figure 7: Reaper induces differential inhibition of translation in an mRNA-dependent manner.

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Acknowledgements

We thank R.M. Elliott, S.S. Margolis, J.E. Irazoqui, K. Silva, T. Prince, A. Vila-Sanjurjo, S. Paranjape, B. Kaplan, E. Harris, R. Rhodes, I.N. Shatsky, E. Nogales and J. Doudna for helpful discussions and generous sharing of advice and reagents. We thank in particular C. Fraser (University of California at Berkeley) for advice and kindly providing purified eIF3 complex and the US National Cell Culture Center for supplying HeLa cells. This work was supported by US National Institutes of Health grants to S.K. (RO1 GM61919) and J.C. (R01 GM65050). R.M. was supported by the Oklahoma Agricultural Experiment Station (Project 1975). C.L.S. was supported by a Howard Hughes Medical Institute Predoctoral Fellowship. D.C.-R. was supported by a US National Institutes of Health Minority Supplement to grant R01 GM61919 and by the Gates Millennium Scholarship. D.C.-R. is a Damon Runyon Fellow.

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Author notes

  1. Daniel A Colón-Ramos

    Present address: Department of Biological Sciences, Stanford University, Stanford, California, 94305, USA

  2. Daniel A Colón-Ramos, Christina L Shenvi, Jamie Cate and Sally Kornbluth: These authors contributed equally to this work.

Authors and Affiliations

  1. Department of Pharmacology and Cancer Biology, Duke University Medical Center, Durham, 27710, North Carolina, USA

    Daniel A Colón-Ramos, Douglas H Weitzel, Eugene C Gan & Sally Kornbluth

  2. Departments of Chemistry and Molecular and Cell Biology, University of California, Berkeley, 94720, California, USA

    Christina L Shenvi & Jamie Cate

  3. Department of Biochemistry and Molecular Biology, Oklahoma State University, Stillwater, 74078, Oklahoma, USA

    Robert Matts

  4. Physical Biosciences Division, Lawrence Berkeley National Laboratory, Berkeley, 94720, California, USA

    Jamie Cate

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Supplementary information

Supplementary Fig. 1

Ribosomes restore Reaper's inhibitory activity but do not enhance translation of control-depleted extracts. (PDF 5728 kb)

Supplementary Fig. 2

Titratable effects of Reaper on polysome sedimentation profiles. (PDF 4561 kb)

Supplementary Fig. 3

Reaper induces accumulation of 80S not associated with the mRNAs. (PDF 5175 kb)

Supplementary Fig. 4

Reaper does not affect phosphorylation of eIF2α or its association with the 40S. (PDF 5121 kb)

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Colón-Ramos, D., Shenvi, C., Weitzel, D. et al. Direct ribosomal binding by a cellular inhibitor of translation. Nat Struct Mol Biol 13, 103–111 (2006). https://doi.org/10.1038/nsmb1052

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