Probing the closed-loop model of mRNA translation in living cells

Stuart K. Archer; Nikolay E. Shirokikh; Claus V. Hallwirth; Traude H. Beilharz; Thomas Preiss

doi:10.1080/15476286.2015.1017242

Probing the closed-loop model of mRNA translation in living cells

Stuart K. Archer, Nikolay E. Shirokikh, Claus V. Hallwirth, Traude H. Beilharz, Thomas Preiss^*

^*Corresponding author for this work

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

45 Citations (Scopus)

Abstract

The mRNA closed-loop, formed through interactions between the cap structure, poly(A) tail, eIF4E, eIF4G and PAB, features centrally in models of eukaryotic translation initiation, although direct support for its existence in vivo is not well established. Here, we investigated the closed-loop using a combination of mRNP isolation from rapidly cross-linked cells and high-throughput qPCR. Using the interaction between these factors and the opposing ends of mRNAs as a proxy for the closed-loop, we provide evidence that it is prevalent for eIF4E/4G-bound but unexpectedly sparse for PAB1-bound mRNAs, suggesting it primarily occurs during a distinct phase of polysome assembly.

Original language	English
Pages (from-to)	248-254
Number of pages	7
Journal	RNA Biology
Volume	12
Issue number	3
DOIs	https://doi.org/10.1080/15476286.2015.1017242
Publication status	Published - 2015

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10.1080/15476286.2015.1017242

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AU - Archer, Stuart K.

AU - Shirokikh, Nikolay E.

AU - Hallwirth, Claus V.

AU - Beilharz, Traude H.

AU - Preiss, Thomas

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AB - The mRNA closed-loop, formed through interactions between the cap structure, poly(A) tail, eIF4E, eIF4G and PAB, features centrally in models of eukaryotic translation initiation, although direct support for its existence in vivo is not well established. Here, we investigated the closed-loop using a combination of mRNP isolation from rapidly cross-linked cells and high-throughput qPCR. Using the interaction between these factors and the opposing ends of mRNAs as a proxy for the closed-loop, we provide evidence that it is prevalent for eIF4E/4G-bound but unexpectedly sparse for PAB1-bound mRNAs, suggesting it primarily occurs during a distinct phase of polysome assembly.

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Probing the closed-loop model of mRNA translation in living cells

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