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Yeast Ded1 promotes 48S translation pre-initiation complex assembly in an mRNA-specific and eIF4F-dependent manner

Neha Gupta, View ORCID ProfileJon R. Lorsch, View ORCID ProfileAlan G. Hinnebusch
doi: https://doi.org/10.1101/353078
Neha Gupta
Eunice Kennedy Shriver National Institute of Child Health and Human Development, National Institutes of Health, Bethesda, MD 20892
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Jon R. Lorsch
Eunice Kennedy Shriver National Institute of Child Health and Human Development, National Institutes of Health, Bethesda, MD 20892
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  • ORCID record for Jon R. Lorsch
  • For correspondence: ahinnebusch@nih.gov jon.lorsch@nih.gov
Alan G. Hinnebusch
Eunice Kennedy Shriver National Institute of Child Health and Human Development, National Institutes of Health, Bethesda, MD 20892
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  • ORCID record for Alan G. Hinnebusch
  • For correspondence: ahinnebusch@nih.gov jon.lorsch@nih.gov
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Summary

DEAD-box RNA helicase Dedl is thought to resolve secondary structures in mRNA 5′-untranslated regions (5′-UTRs) that impede 48S preinitiation complex (PIC) formation at the initiation codon. We reconstituted Ded1 acceleration of 48S PIC assembly on native mRNAs in a pure system, and recapitulated increased Ded1-dependence of mRNAs that are Ded1-hyperdependent in vivo. Stem-loop (SL) structures in 5′-UTRs of native and synthetic mRNAs increased the Ded1 requirement to overcome their intrinsically low rates of 48S PIC recruitment. Ded1 acceleration of 48S assembly was greater in the presence of eIF4F, and domains mediating one or more Ded1 interactions with eIF4G or helicase eIF4A were required for efficient recruitment of all mRNAs; however, the relative importance of particular Ded1 and eIF4G domains were distinct for each mRNA. Our results account for the Ded1 hyper-dependence of mRNAs with structure-prone 5′-UTRs, and implicate an eIF4E·eIF4G·eIF4A·Ded1 complex in accelerating 48S PIC assembly on native mRNAs.

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The copyright holder for this preprint is the author/funder, who has granted bioRxiv a license to display the preprint in perpetuity. It is made available under a CC-BY 4.0 International license.
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Posted June 21, 2018.
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Yeast Ded1 promotes 48S translation pre-initiation complex assembly in an mRNA-specific and eIF4F-dependent manner
Neha Gupta, Jon R. Lorsch, Alan G. Hinnebusch
bioRxiv 353078; doi: https://doi.org/10.1101/353078
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Yeast Ded1 promotes 48S translation pre-initiation complex assembly in an mRNA-specific and eIF4F-dependent manner
Neha Gupta, Jon R. Lorsch, Alan G. Hinnebusch
bioRxiv 353078; doi: https://doi.org/10.1101/353078

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