Interaction between eukaryotic initiation factors 1A and 5B is required for efficient ribosomal subunit joining

Michael G. Acker, Byung Sik Shin, Thomas E. Dever, Jon R. Lorsch

Research output: Contribution to journalArticle

Abstract

Eukaryotic initiation factor 5B (eIF5B) is a GTPase that facilitates joining of the 60 S ribosomal subunit to the 40 S ribosomal subunit during translation initiation. Formation of the resulting 80 S initiation complex triggers eIF5B to hydrolyze its bound GTP, reducing the affinity of the factor for the complex and allowing it to dissociate. Here we present a kinetic analysis of GTP hydrolysis by eIF5B in the context of the translation initiation pathway. Our data indicate that stimulation of GTP hydrolysis by eIF5B requires the completion of early steps in translation initiation, including the eIF1- and eIF1A-dependent delivery of initiator methionyl-tRNA to the 40 S ribosomal subunit and subsequent GTP hydrolysis by eIF2. Full activation of GTP hydrolysis by eIF5B requires the extreme C terminus of eIF1A, which has previously been shown to interact with the C terminus of eIF5B. Disruption of either isoleucine residue in the eIF1A C-terminal sequence DIDDI reduces the rate constant for GTP hydrolysis by ∼20-fold, whereas changing the aspartic acid residues has no effect. Changing the isoleucines in the C terminus of eIF1A also disrupts the ability of eIF5B to facilitate subunit joining. These data indicate that the interaction of the C terminus of eIF1A with eIF5B promotes ribosomal subunit joining and possibly provides a checkpoint for correct complex formation, allowing full activation of GTP hydrolysis only upon formation of a properly organized 80 S initiation complex.

Original languageEnglish (US)
Pages (from-to)8469-8475
Number of pages7
JournalJournal of Biological Chemistry
Volume281
Issue number13
DOIs
StatePublished - Mar 31 2006

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Eukaryotic Initiation Factors
Ribosome Subunits
Joining
Guanosine Triphosphate
Hydrolysis
Isoleucine
Chemical activation
RNA, Transfer, Met
eukaryotic initiation factor-5B
GTP Phosphohydrolases
Transfer RNA
Aspartic Acid
Rate constants
Kinetics

ASJC Scopus subject areas

  • Biochemistry

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Interaction between eukaryotic initiation factors 1A and 5B is required for efficient ribosomal subunit joining. / Acker, Michael G.; Shin, Byung Sik; Dever, Thomas E.; Lorsch, Jon R.

In: Journal of Biological Chemistry, Vol. 281, No. 13, 31.03.2006, p. 8469-8475.

Research output: Contribution to journalArticle

Acker, Michael G. ; Shin, Byung Sik ; Dever, Thomas E. ; Lorsch, Jon R. / Interaction between eukaryotic initiation factors 1A and 5B is required for efficient ribosomal subunit joining. In: Journal of Biological Chemistry. 2006 ; Vol. 281, No. 13. pp. 8469-8475.
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