Yeast Ded1 promotes 48S translation pre-initiation complex assembly in an mRNA-specific and eIF4F-dependent manner
DEAD-box RNA helicase Ded1 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 recapi...
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doaj-f9b3b39aa6e64d49b72b64614ae651182021-05-05T16:11:18ZengeLife Sciences Publications LtdeLife2050-084X2018-10-01710.7554/eLife.38892Yeast Ded1 promotes 48S translation pre-initiation complex assembly in an mRNA-specific and eIF4F-dependent mannerNeha Gupta0https://orcid.org/0000-0002-1845-2940Jon R Lorsch1https://orcid.org/0000-0002-4521-4999Alan G Hinnebusch2https://orcid.org/0000-0002-1627-8395Eunice Kennedy Shriver National Institute of Child Health and Human Development, National Institutes of Health, Bethesda, United StatesEunice Kennedy Shriver National Institute of Child Health and Human Development, National Institutes of Health, Bethesda, United StatesEunice Kennedy Shriver National Institute of Child Health and Human Development, National Institutes of Health, Bethesda, United StatesDEAD-box RNA helicase Ded1 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.https://elifesciences.org/articles/38892translationinitiationDed1eIF4Gyeast |
collection |
DOAJ |
language |
English |
format |
Article |
sources |
DOAJ |
author |
Neha Gupta Jon R Lorsch Alan G Hinnebusch |
spellingShingle |
Neha Gupta Jon R Lorsch Alan G Hinnebusch Yeast Ded1 promotes 48S translation pre-initiation complex assembly in an mRNA-specific and eIF4F-dependent manner eLife translation initiation Ded1 eIF4G yeast |
author_facet |
Neha Gupta Jon R Lorsch Alan G Hinnebusch |
author_sort |
Neha Gupta |
title |
Yeast Ded1 promotes 48S translation pre-initiation complex assembly in an mRNA-specific and eIF4F-dependent manner |
title_short |
Yeast Ded1 promotes 48S translation pre-initiation complex assembly in an mRNA-specific and eIF4F-dependent manner |
title_full |
Yeast Ded1 promotes 48S translation pre-initiation complex assembly in an mRNA-specific and eIF4F-dependent manner |
title_fullStr |
Yeast Ded1 promotes 48S translation pre-initiation complex assembly in an mRNA-specific and eIF4F-dependent manner |
title_full_unstemmed |
Yeast Ded1 promotes 48S translation pre-initiation complex assembly in an mRNA-specific and eIF4F-dependent manner |
title_sort |
yeast ded1 promotes 48s translation pre-initiation complex assembly in an mrna-specific and eif4f-dependent manner |
publisher |
eLife Sciences Publications Ltd |
series |
eLife |
issn |
2050-084X |
publishDate |
2018-10-01 |
description |
DEAD-box RNA helicase Ded1 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. |
topic |
translation initiation Ded1 eIF4G yeast |
url |
https://elifesciences.org/articles/38892 |
work_keys_str_mv |
AT nehagupta yeastded1promotes48stranslationpreinitiationcomplexassemblyinanmrnaspecificandeif4fdependentmanner AT jonrlorsch yeastded1promotes48stranslationpreinitiationcomplexassemblyinanmrnaspecificandeif4fdependentmanner AT alanghinnebusch yeastded1promotes48stranslationpreinitiationcomplexassemblyinanmrnaspecificandeif4fdependentmanner |
_version_ |
1721459620737712128 |