Eukaryotic translation initiation factor eIF5 promotes the accuracy of start codon recognition by regulating Pi release and conformational transitions of the preinitiation complex

Adesh K. Saini, Jagpreet S. Nanda, Pilar Martin-Marcos, Jinsheng Dong, Fan Zhang, Monika Bhardwaj, Jon R. Lorsch, Alan G. Hinnebusch

Research output: Contribution to journalArticle

Abstract

eIF5 is the GTPase activating protein (GAP) for the eIF2 · GTP · Met-tRNAi (Met) ternary complex with a critical role in initiation codon selection. Previous work suggested that the eIF5 mutation G31R/SUI5 elevates initiation at UUG codons by increasing GAP function. Subsequent work implicated eIF5 in rearrangement of the preinitiation complex (PIC) from an open, scanning conformation to a closed state at AUG codons, from which Pi is released from eIF2 · GDP · Pi. To identify eIF5 functions crucial for accurate initiation, we investigated the consequences of G31R on GTP hydrolysis and Pi release, and the effects of intragenic G31R suppressors on these reactions, and on the partitioning of PICs between open and closed states. eIF5-G31R altered regulation of Pi release, accelerating it at UUG while decreasing it at AUG codons, consistent with its ability to stabilize the closed complex at UUG. Suppressor G62S mitigates both defects of G31R, accounting for its efficient suppression of UUG initiation in G31R,G62S cells; however suppressor M18V impairs GTP hydrolysis with little effect on PIC conformation. The strong defect in GTP hydrolysis conferred by M18V likely explains its broad suppression of Sui(-) mutations in numerous factors. We conclude that both of eIF5's functions, regulating Pi release and stabilizing the closed PIC conformation, contribute to stringent AUG selection in vivo.

Original languageEnglish (US)
Pages (from-to)9623-9640
Number of pages18
JournalNucleic Acids Research
Volume42
Issue number15
DOIs
StatePublished - Sep 1 2014
Externally publishedYes

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Eukaryotic Initiation Factors
Initiator Codon
Guanosine Triphosphate
Codon
GTPase-Activating Proteins
Hydrolysis
Mutation

ASJC Scopus subject areas

  • Medicine(all)

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Eukaryotic translation initiation factor eIF5 promotes the accuracy of start codon recognition by regulating Pi release and conformational transitions of the preinitiation complex. / Saini, Adesh K.; Nanda, Jagpreet S.; Martin-Marcos, Pilar; Dong, Jinsheng; Zhang, Fan; Bhardwaj, Monika; Lorsch, Jon R.; Hinnebusch, Alan G.

In: Nucleic Acids Research, Vol. 42, No. 15, 01.09.2014, p. 9623-9640.

Research output: Contribution to journalArticle

Saini, Adesh K. ; Nanda, Jagpreet S. ; Martin-Marcos, Pilar ; Dong, Jinsheng ; Zhang, Fan ; Bhardwaj, Monika ; Lorsch, Jon R. ; Hinnebusch, Alan G. / Eukaryotic translation initiation factor eIF5 promotes the accuracy of start codon recognition by regulating Pi release and conformational transitions of the preinitiation complex. In: Nucleic Acids Research. 2014 ; Vol. 42, No. 15. pp. 9623-9640.
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