Crystal Structure of the C-terminal Domain of S.cerevisiae eIF5

Wei, Zhiyi; Xue, Yongjun; Xu, Hang; Gong, Weimin

doi:10.1016/j.jmb.2006.03.037

Cited by 40 publications

(43 citation statements)

References 38 publications

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“…The equivalent eIF5-7A mutation had similar effects on function and eIF5/ eIF2␤ interaction, leading to the idea that GEF and GTPaseactivating protein share a common interaction site on eIF2␤ (6). The crystal structures of both ε cat and eIF5-CTD confirm that they share a highly similar helical HEAT repeat structure and that the εW699 (W391 in yeast eIF5) side chain is exposed on the surface at one end of the protein, predicting that it would be important for eIF2 interaction (8,9,40) (Fig. 1B).…”

Section: Discussionmentioning

confidence: 99%

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Critical Contacts between the Eukaryotic Initiation Factor 2B (eIF2B) Catalytic Domain and both eIF2β and -2γ Mediate Guanine Nucleotide Exchange

Mohammad-Qureshi

Haddad

Hemingway

et al. 2007

Molecular and Cellular Biology

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Diverse guanine nucleotide exchange factors (GEFs) regulate the activity of GTP binding proteins. One of the most complicated pairs is eukaryotic initiation factor 2B (eIF2B) and eIF2, which function during protein synthesis initiation in eukaryotes. We have mutated conserved surface residues within the eIF2B GEF domain, located at the eIF2B C terminus. Extensive genetic and biochemical characterization established how these residues contribute to GEF activity. We find that the universally conserved residue E569 is critical for activity and that even a conservative E569D substitution is lethal in vivo. Several mutations within residues close to E569 have no discernible effect on growth or GCN4 expression, but an alanine substitution at the adjacent L568 is cold sensitive and deregulates GCN4 activity at 15°C. The mutation of W699, found on a separate surface approximately 40 Å from E569, is also lethal. Binding studies show that W699 is critical for interaction with eIF2␤, while L568 and E569 are not. In contrast, all three residues are critical for interaction with eIF2␥. These data show that multiple contacts between eIF2␥ and eIF2B mediate nucleotide exchange.The cap-dependent pathway for the initiation of translation requires the assembly of eukaryotic initiation factor (eIF) ribosomal subunits and a selected mRNA. Central to the initiation process is the GTP binding protein eIF2, which delivers aminoacylated initiator methionyl tRNA (Met-tRNA i Met ) to the 40S ribosome as part of a multifactor complex containing eIFs 1, 3, and 5 (5). This 43S preinitiation complex associates with the 5Ј end of an mRNA and migrates along it to locate an AUG initiator codon with the aid of other factors. AUG codon recognition, eIF5-promoted hydrolysis of GTP bound to eIF2, and phosphate release stimulate eIF2-GDP and eIF5 dissociation from the initiation complex, probably as an eIF2-GDP/ eIF5 complex (33). Met-tRNA i Met remains bound to the 40S ribosomal subunit at the AUG and is probably stabilized by eIF1A/eIF5B-GTP. 60S ribosomal subunit joining is accelerated by GTP hydrolysis and the release of eIF5B-GDP. At this point, translation elongation can commence (20).The regeneration of eIF2-GTP from the inactive GDPbound complex released from the initiation complex is carried out by the guanine nucleotide exchange factor (GEF) eIF2B. eIF2B accelerates the otherwise slow dissociation of GDP from eIF2, allowing its replacement with GTP. This process is regulated by the phosphorylation of eIF2␣ on the conserved ser51 residue in response to a diverse set of cellular stresses (20). For example, in yeast cells (Saccharomyces cerevisiae), translational control of GCN4 expression proceeds by the following pathway: amino acid starvation activates Gcn2p, which phosphorylates eIF2␣ and inhibits eIF2B activity. This lowers the rate of nucleotide exchange and slows recruitment of the eIF2-GTP/Met-tRNA i Met ternary complex (TC) to mRNAs. GCN4 translation is activated by this response, as it contains short upstream open reading frames tha...

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Section: Discussionmentioning

confidence: 99%

“…The functional significance of these sequence similarities is not clear. In addition, the C-terminal domain (CTD) of eIF2Bε is homologous both to the eIF5-CTD and to eIF4G (pfam02020) (8,9,40). It is this last domain that contains residues sufficient for GEF activity of eIF2B, albeit at a reduced rate compared with that of the full five-subunit complex (21).…”

mentioning

confidence: 99%

Critical Contacts between the Eukaryotic Initiation Factor 2B (eIF2B) Catalytic Domain and both eIF2β and -2γ Mediate Guanine Nucleotide Exchange

Mohammad-Qureshi

Haddad

Hemingway

et al. 2007

Molecular and Cellular Biology

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“…eIF5 has two functional domains, N-terminal (NTD) and C-terminal (CTD) with a linker connection (Conte et al, 2006;Wei et al, 2006). An "arginine finger" (Arg-15) required for GTPase activity is positioned near the N-terminus in an unstructured region.…”

Section: Eif5mentioning

confidence: 99%

Mechanism of Cytoplasmic mRNA Translation

Browning

Bailey‐Serres

2015

The Arabidopsis Book

187

220

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Protein synthesis is a fundamental process in gene expression that depends upon the abundance and accessibility of the mRNA transcript as well as the activity of many protein and RNA-protein complexes. Here we focus on the intricate mechanics of mRNA translation in the cytoplasm of higher plants. This chapter includes an inventory of the plant translational apparatus and a detailed review of the translational processes of initiation, elongation, and termination. The majority of mechanistic studies of cytoplasmic translation have been carried out in yeast and mammalian systems. The factors and mechanisms of translation are for the most part conserved across eukaryotes; however, some distinctions are known to exist in plants. A comprehensive understanding of the complex translational apparatus and its regulation in plants is warranted, as the modulation of protein production is critical to development, environmental plasticity and biomass yield in diverse ecosystems and agricultural settings.

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“…The eIF2a residue Ser51 (blue), GTP analog (green), Met-tRNA i Met (gray), and mRNA (cyan) with AUG codon (yellow) are indicated. (B) eIF5 domains and activities (left) and structural models (right) for the human GAP domain bearing R15 (pdb 2E9H) and the yeast CTD bearing W391 (pdb 2FUL) (Wei et al 2006). Structures were drawn using Chimera software (University of California, San Francisco, UCSF).…”

Section: Ternary Complex Formationmentioning

confidence: 99%

Mechanism and Regulation of Protein Synthesis in Saccharomyces cerevisiae

2016

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In this review, we provide an overview of protein synthesis in the yeast Saccharomyces cerevisiae. The mechanism of protein synthesis is well conserved between yeast and other eukaryotes, and molecular genetic studies in budding yeast have provided critical insights into the fundamental process of translation as well as its regulation. The review focuses on the initiation and elongation phases of protein synthesis with descriptions of the roles of translation initiation and elongation factors that assist the ribosome in binding the messenger RNA (mRNA), selecting the start codon, and synthesizing the polypeptide. We also examine mechanisms of translational control highlighting the mRNA cap-binding proteins and the regulation of GCN4 and CPA1 mRNAs.

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Crystal Structure of the C-terminal Domain of S.cerevisiae eIF5

Cited by 40 publications

References 38 publications

Critical Contacts between the Eukaryotic Initiation Factor 2B (eIF2B) Catalytic Domain and both eIF2β and -2γ Mediate Guanine Nucleotide Exchange

Critical Contacts between the Eukaryotic Initiation Factor 2B (eIF2B) Catalytic Domain and both eIF2β and -2γ Mediate Guanine Nucleotide Exchange

Mechanism of Cytoplasmic mRNA Translation

Mechanism and Regulation of Protein Synthesis in Saccharomyces cerevisiae

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