Archaeal aIF2B Interacts with Eukaryotic Translation Initiation Factors eIF2α and eIF2Bα: Implications for aIF2B Function and eIF2B Regulation

Kamal Dev, Thomas J. Santangelo, Stefan Rothenburg, Dante Neculai, Madhusudan Dey, Frank Sicheri, Thomas E. Dever, John N. Reeve, Alan G. Hinnebusch

Research output: Contribution to journalArticle

22 Citations (Scopus)

Abstract

Translation initiation is down-regulated in eukaryotes by phosphorylation of the α-subunit of eIF2 (eukaryotic initiation factor 2), which inhibits its guanine nucleotide exchange factor, eIF2B. The N-terminal S1 domain of phosphorylated eIF2α interacts with a subcomplex of eIF2B formed by the three regulatory subunits α/GCN3, β/GCD7, and δ/GCD2, blocking the GDP-GTP exchange activity of the catalytic e{open}-subunit of eIF2B. These regulatory subunits have related sequences and have sequences in common with many archaeal proteins, some of which are involved in methionine salvage and CO2 fixation. Our sequence analyses however predicted that members of one phylogenetically distinct and coherent group of these archaeal proteins [designated aIF2Bs (archaeal initiation factor 2Bs)] are functional homologs of the α, β, and δ subunits of eIF2B. Three of these proteins, from different archaea, have been shown to bind in vitro to the α-subunit of the archaeal aIF2 from the cognate archaeon. In one case, the aIF2B protein was shown further to bind to the S1 domain of the α-subunit of yeast eIF2 in vitro and to interact with eIF2Bα/GCN3 in vivo in yeast. The aIF2B-eIF2α interaction was however independent of eIF2α phosphorylation. Mass spectrometry has identified several proteins that co-purify with aIF2B from Thermococcus kodakaraensis, and these include aIF2α, a sugar-phosphate nucleotidyltransferase with sequence similarity to eIF2Be{open}, and several large-subunit (50S) ribosomal proteins. Based on this evidence that aIF2B has functions in common with eIF2B, the crystal structure established for an aIF2B was used to construct a model of the eIF2B regulatory subcomplex. In this model, the evolutionarily conserved regions and sites of regulatory mutations in the three eIF2B subunits in yeast are juxtaposed in one continuous binding surface for phosphorylated eIF2α.

Original languageEnglish (US)
Pages (from-to)701-722
Number of pages22
JournalJournal of Molecular Biology
Volume392
Issue number3
DOIs
StatePublished - Sep 25 2009
Externally publishedYes

Fingerprint

Eukaryotic Initiation Factor-2
Eukaryotic Initiation Factors
Archaeal Proteins
Yeasts
Archaea
Nucleotidyltransferases
Thermococcus
Phosphorylation
Sugar Phosphates
Guanine Nucleotide Exchange Factors
Peptide Initiation Factors
Proteins
Ribosomal Proteins
Nucleic Acid Regulatory Sequences
Guanosine Triphosphate
Eukaryota
Methionine
Sequence Analysis
Mass Spectrometry
Mutation

Keywords

  • aIF2B
  • archaea
  • eIF2
  • eIF2B
  • S. cerevisiae

ASJC Scopus subject areas

  • Molecular Biology

Cite this

Archaeal aIF2B Interacts with Eukaryotic Translation Initiation Factors eIF2α and eIF2Bα : Implications for aIF2B Function and eIF2B Regulation. / Dev, Kamal; Santangelo, Thomas J.; Rothenburg, Stefan; Neculai, Dante; Dey, Madhusudan; Sicheri, Frank; Dever, Thomas E.; Reeve, John N.; Hinnebusch, Alan G.

In: Journal of Molecular Biology, Vol. 392, No. 3, 25.09.2009, p. 701-722.

Research output: Contribution to journalArticle

Dev, Kamal ; Santangelo, Thomas J. ; Rothenburg, Stefan ; Neculai, Dante ; Dey, Madhusudan ; Sicheri, Frank ; Dever, Thomas E. ; Reeve, John N. ; Hinnebusch, Alan G. / Archaeal aIF2B Interacts with Eukaryotic Translation Initiation Factors eIF2α and eIF2Bα : Implications for aIF2B Function and eIF2B Regulation. In: Journal of Molecular Biology. 2009 ; Vol. 392, No. 3. pp. 701-722.
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abstract = "Translation initiation is down-regulated in eukaryotes by phosphorylation of the α-subunit of eIF2 (eukaryotic initiation factor 2), which inhibits its guanine nucleotide exchange factor, eIF2B. The N-terminal S1 domain of phosphorylated eIF2α interacts with a subcomplex of eIF2B formed by the three regulatory subunits α/GCN3, β/GCD7, and δ/GCD2, blocking the GDP-GTP exchange activity of the catalytic e{open}-subunit of eIF2B. These regulatory subunits have related sequences and have sequences in common with many archaeal proteins, some of which are involved in methionine salvage and CO2 fixation. Our sequence analyses however predicted that members of one phylogenetically distinct and coherent group of these archaeal proteins [designated aIF2Bs (archaeal initiation factor 2Bs)] are functional homologs of the α, β, and δ subunits of eIF2B. Three of these proteins, from different archaea, have been shown to bind in vitro to the α-subunit of the archaeal aIF2 from the cognate archaeon. In one case, the aIF2B protein was shown further to bind to the S1 domain of the α-subunit of yeast eIF2 in vitro and to interact with eIF2Bα/GCN3 in vivo in yeast. The aIF2B-eIF2α interaction was however independent of eIF2α phosphorylation. Mass spectrometry has identified several proteins that co-purify with aIF2B from Thermococcus kodakaraensis, and these include aIF2α, a sugar-phosphate nucleotidyltransferase with sequence similarity to eIF2Be{open}, and several large-subunit (50S) ribosomal proteins. Based on this evidence that aIF2B has functions in common with eIF2B, the crystal structure established for an aIF2B was used to construct a model of the eIF2B regulatory subcomplex. In this model, the evolutionarily conserved regions and sites of regulatory mutations in the three eIF2B subunits in yeast are juxtaposed in one continuous binding surface for phosphorylated eIF2α.",
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