Reinitiation and other unconventional posttermination events during eukaryotic translation

Maxim A. Skabkin; Olga V. Skabkina; Christopher U.T. Hellen; Tatyana V. Pestova

doi:10.1016/j.molcel.2013.05.026

Reinitiation and other unconventional posttermination events during eukaryotic translation

Maxim A. Skabkin
, Olga V. Skabkina
, Christopher U.T. Hellen
, Tatyana V. Pestova

Cell / Cellular Biology and Anatomical Sciences

SUNY Downstate Health Sciences University

SUNY Downstate Health Sciences University

Research output: Contribution to journal › Article › peer-review

127 Scopus citations

Abstract

During ribosome recycling, posttermination complexes are dissociated by ABCE1 and eRF1 into 60Sand tRNA/mRNA-associated 40S subunits, after which tRNA and mRNA are released by eIF1/eIF1A, Ligatin, or MCT-1/DENR. Occasionally, 40S subunits remain associated with mRNA and reinitiate at nearby AUGs. We recapitulated reinitiation using a reconstituted mammalian translation system. The presence of eIF2, eIF3, eIF1, eIF1A, and Met-tRNA_i^Met was sufficient for recycled 40S subunits to remain on mRNA, scan bidirectionally, and reinitiate at upstream and downstream AUGs if mRNA regions flanking the stop codon were unstructured. Imposition of3' directionality additionally required eIF4F. Strikingly, posttermination ribosomes were not stably anchored on mRNA and migrated bidirectionally tocodons cognate to the P site tRNA. Migration depended onthe mode of peptide release (puromycin> eRF1{dot operator}eRF3) and nature of tRNA and was enhanced by eEF2. The mobility of postterminationribosomes suggests that some reinitiation eventscould involve 80S ribosomes rather than 40S subunits.

Original language	English
Pages (from-to)	249-264
Number of pages	16
Journal	Molecular Cell
Volume	51
Issue number	2
DOIs	https://doi.org/10.1016/j.molcel.2013.05.026
State	Published - Jul 25 2013

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title = "Reinitiation and other unconventional posttermination events during eukaryotic translation",

abstract = "During ribosome recycling, posttermination complexes are dissociated by ABCE1 and eRF1 into 60Sand tRNA/mRNA-associated 40S subunits, after which tRNA and mRNA are released by eIF1/eIF1A, Ligatin, or MCT-1/DENR. Occasionally, 40S subunits remain associated with mRNA and reinitiate at nearby AUGs. We recapitulated reinitiation using a reconstituted mammalian translation system. The presence of eIF2, eIF3, eIF1, eIF1A, and Met-tRNAiMet was sufficient for recycled 40S subunits to remain on mRNA, scan bidirectionally, and reinitiate at upstream and downstream AUGs if mRNA regions flanking the stop codon were unstructured. Imposition of3' directionality additionally required eIF4F. Strikingly, posttermination ribosomes were not stably anchored on mRNA and migrated bidirectionally tocodons cognate to the P site tRNA. Migration depended onthe mode of peptide release (puromycin> eRF1\{dot operator\}eRF3) and nature of tRNA and was enhanced by eEF2. The mobility of postterminationribosomes suggests that some reinitiation eventscould involve 80S ribosomes rather than 40S subunits.",

author = "Skabkin, \{Maxim A.\} and Skabkina, \{Olga V.\} and Hellen, \{Christopher U.T.\} and Pestova, \{Tatyana V.\}",

year = "2013",

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doi = "10.1016/j.molcel.2013.05.026",

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T1 - Reinitiation and other unconventional posttermination events during eukaryotic translation

AU - Skabkin, Maxim A.

AU - Skabkina, Olga V.

AU - Hellen, Christopher U.T.

AU - Pestova, Tatyana V.

PY - 2013/7/25

Y1 - 2013/7/25

N2 - During ribosome recycling, posttermination complexes are dissociated by ABCE1 and eRF1 into 60Sand tRNA/mRNA-associated 40S subunits, after which tRNA and mRNA are released by eIF1/eIF1A, Ligatin, or MCT-1/DENR. Occasionally, 40S subunits remain associated with mRNA and reinitiate at nearby AUGs. We recapitulated reinitiation using a reconstituted mammalian translation system. The presence of eIF2, eIF3, eIF1, eIF1A, and Met-tRNAiMet was sufficient for recycled 40S subunits to remain on mRNA, scan bidirectionally, and reinitiate at upstream and downstream AUGs if mRNA regions flanking the stop codon were unstructured. Imposition of3' directionality additionally required eIF4F. Strikingly, posttermination ribosomes were not stably anchored on mRNA and migrated bidirectionally tocodons cognate to the P site tRNA. Migration depended onthe mode of peptide release (puromycin> eRF1{dot operator}eRF3) and nature of tRNA and was enhanced by eEF2. The mobility of postterminationribosomes suggests that some reinitiation eventscould involve 80S ribosomes rather than 40S subunits.

AB - During ribosome recycling, posttermination complexes are dissociated by ABCE1 and eRF1 into 60Sand tRNA/mRNA-associated 40S subunits, after which tRNA and mRNA are released by eIF1/eIF1A, Ligatin, or MCT-1/DENR. Occasionally, 40S subunits remain associated with mRNA and reinitiate at nearby AUGs. We recapitulated reinitiation using a reconstituted mammalian translation system. The presence of eIF2, eIF3, eIF1, eIF1A, and Met-tRNAiMet was sufficient for recycled 40S subunits to remain on mRNA, scan bidirectionally, and reinitiate at upstream and downstream AUGs if mRNA regions flanking the stop codon were unstructured. Imposition of3' directionality additionally required eIF4F. Strikingly, posttermination ribosomes were not stably anchored on mRNA and migrated bidirectionally tocodons cognate to the P site tRNA. Migration depended onthe mode of peptide release (puromycin> eRF1{dot operator}eRF3) and nature of tRNA and was enhanced by eEF2. The mobility of postterminationribosomes suggests that some reinitiation eventscould involve 80S ribosomes rather than 40S subunits.

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U2 - 10.1016/j.molcel.2013.05.026

DO - 10.1016/j.molcel.2013.05.026

M3 - Article

C2 - 23810859

SN - 1097-2765

VL - 51

SP - 249

EP - 264

JO - Molecular Cell

JF - Molecular Cell

IS - 2

ER -

Reinitiation and other unconventional posttermination events during eukaryotic translation

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