Crystal Structure and Functional Analysis of the Eukaryotic Class II Release Factor eRF3 from S. pombe

Kong, Chunguang; Ito, Koichi; Walsh, Martin A.; Wada, Miki; Liu, Yuying; Kumar, Sushil; Barford, David; Nakamura, Yoshikazu; Song, Haiwei

doi:10.1016/s1097-2765(04)00206-0

Cited by 118 publications

(169 citation statements)

References 54 publications

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Section: Resultsmentioning

confidence: 99%

“…In this region, the highly conserved GRFTLRD motif plays a crucial role in mediating eRF1 binding (Kong et al 2004). eRF3 variants with either substitutions in the GRFTLRD motif or C-terminal truncation show reduced interaction with eRF1 (Kong et al 2004). To examine the influence of eRF1 binding disruption on eRF3a stability, human eRF3a variants carrying either a deletion of the last 31 C-terminal amino acids (eRF3a-DC) or mutations in the GRFTLRD motif (eRF3a-FRAA) were created and overexpressed in HEK293 cells.…”

Section: Resultsmentioning

confidence: 99%

“…Thus, it is postulated that, in eukaryotic cells, eRF3 is predominantly bound to GTP in a relatively long-lived eRF1deRF3dGTP ternary complex, and eRF3 that is not associated with eRF1 is likely to be in the GDP-bound form (Hauryliuk et al 2006;Pisareva et al 2006). These studies, together with the data obtained from the crystal structure of yeast eRF3 (Kong et al 2004), indicate that the mechanism of nucleotide binding/exchange on eRF3 is very distinct among GTPbinding proteins: Mg 2+ does not stabilize GDP binding to eRF3 alone or to its complex with eRF1, and a GTP/GDP exchange factor (GEF) is not required for rapid GDP dissociation from any form of the factor (Kong et al 2004;Pisareva et al 2006).…”

Section: Introductionmentioning

confidence: 87%

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Proteasomal degradation of human release factor eRF3a regulates translation termination complex formation

Chauvin¹,

Jean-Jean²

2007

RNA

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In eukaryotes, eRF1 and eRF3 are associated in a complex that mediates translation termination. The regulation of the formation of this complex in vivo is far from being understood. In mammalian cells, depletion of eRF3a causes a reduction of eRF1 level by decreasing its stability. Here, we investigate the status of eRF3a when not associated with eRF1. We show that eRF3a forms altered in their eRF1-binding site have a decreased stability, which increases upon cell treatment with the proteasome inhibitor MG132. We also show that eRF3a forms altered in eRF1 binding as well as wild-type eRF3a are polyubiquitinated. These results indicate that eRF3a is degraded by the proteasome when not associated with eRF1 and suggest that proteasomal degradation of eRF3a controls translation termination complex formation by adjusting the eRF3a level to that of eRF1.

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Section: Resultsmentioning

confidence: 99%

Section: Resultsmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 87%

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Proteasomal degradation of human release factor eRF3a regulates translation termination complex formation

Chauvin¹,

Jean-Jean²

2007

RNA

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“…The Ure2 system is therefore a useful model not only to investigate the prion concept, but also to understand the properties of Gln/ Asn-repeat proteins and hence the molecular basis of the related diseases. [27,29,58], Sup35 [114,115], Rnq1 [13], Het-s [22] and PrP [23,24,116,117]. The functional regions are as indicated.…”

Section: Introductionmentioning

confidence: 99%

The yeast prion protein Ure2: Structure, function and folding

Lian

Jiang

Zhang

et al. 2006

Biochimica et Biophysica Acta (BBA) - Proteins and Proteomics

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The Saccharomyces cerevisiae protein Ure2 functions as a regulator of nitrogen metabolism and as a glutathione-dependent peroxidase. Ure2 also has the characteristics of a prion, in that it can undergo a heritable conformational change to an aggregated state; the prion form of Ure2 loses the regulatory function, but the enzymatic function appears to be maintained. A number of factors are found to affect the prion properties of Ure2, including mutation and expression levels of molecular chaperones, and the effect of these factors on structure and stability are being investigated. The relationship between structure, function and folding for the yeast prion Ure2 are discussed.

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“…eRF3 consists of an N-terminal sequence (residues 1-138) that is nonessential for termination (15), and the following essential region comprising a G domain and the β-barrel domains 2 and 3. These three domains share strong structural homology with elongation factors (EFs) eEF1α and EF-Tu (16). Domain 3 interacts directly with domain C of eRF1, predominantly through hydrophobic contacts (17).…”

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confidence: 99%

Cryo-EM structure of the mammalian eukaryotic release factor eRF1–eRF3-associated termination complex

Taylor

Unbehaun

et al. 2012

Proc. Natl. Acad. Sci. U.S.A.

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Eukaryotic translation termination results from the complex functional interplay between two eukaryotic release factors, eRF1 and eRF3, and the ribosome, in which GTP hydrolysis by eRF3 couples codon recognition with peptidyl-tRNA hydrolysis by eRF1. Here, using cryo-electron microscopy (cryo-EM) and flexible fitting, we determined the structure of eRF1-eRF3-guanosine 5′- [β,γ-imido] triphosphate (GMPPNP)-bound ribosomal pretermination complex (pre-TC), which corresponds to the initial, pre-GTP hydrolysis stage of factor attachment. Our results show that eukaryotic translation termination involves a network of interactions between the two release factors and the ribosome. Our structure provides mechanistic insight into the coordination between GTP hydrolysis by eRF3 and subsequent peptide release by eRF1.T ermination of translation occurs when a ribosome reaches the end of the coding region and a stop codon (UAA, UAG, or UGA) enters the aminoacyl tRNA binding site (A site), leaving peptidyl-tRNA in the peptidyl tRNA binding site (P site). It entails stop codon recognition by specialized release factors followed by hydrolysis of peptidyl-tRNA. In eukaryotes, termination is mediated by the concerted action of two directly interacting release factors, eRF1 and eRF3. eRF1 is responsible for stop codon recognition and inducing hydrolysis of peptidyl-tRNA, whereas eRF3, a ribosome-dependent GTPase, strongly stimulates peptide release by eRF1 in a GTP-dependent manner (for review, see ref. 1).eRF1 also participates in ribosome recycling: after peptide release it remains associated with the ribosome and together with ATP-binding cassette sub-family E member 1 (ABCE1) promotes splitting the ribosome into free 60S and tRNA/mRNA-associated 40S subunits (2). eRF1 and eRF3 have paralogs, Dom34 (yeast)/ Pelota (mammals) and Hbs1, respectively, which do not participate in termination but instead play a key role in No-go and nonstop decay surveillance mechanisms (e.g., see refs. 3, 4). Dom34/Pelota and Hbs1 do not induce peptide release in a mechanism similar to eRFs. Instead, they cooperate with ABCE1 to promote dissociation of stalled elongation complexes, which is accompanied by peptidyltRNA drop off (5-7). eRF1 comprises N-terminal (N), middle (M), and C-terminal (C) domains (8). The N domain is responsible for stop codon recognition, which is achieved through a 3D network of conserved residues that include apical TASNIKS (amino acid sequence: threonine-alanine-serine-asparagine-isoleucine-lysine-serine) and YxCxxxF motifs (e.g., refs. 9-12). Domain M contains the universally conserved GGQ motif, which is critical for triggering peptide release: as shown for prokaryotes, its placement into the peptidyl transferase center (PTC) causes rRNA rearrangement, allowing a water molecule to enter (for review, see ref. 13). The rigid core of domain C forms an α-β sandwich (8) that deviates from the standard form by the presence of a small insertion, which forms a minidomain (14). eRF3 consists of an N-terminal sequence (residues 1-...

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Crystal Structure and Functional Analysis of the Eukaryotic Class II Release Factor eRF3 from S. pombe

Cited by 118 publications

References 54 publications

Proteasomal degradation of human release factor eRF3a regulates translation termination complex formation

Proteasomal degradation of human release factor eRF3a regulates translation termination complex formation

The yeast prion protein Ure2: Structure, function and folding

Cryo-EM structure of the mammalian eukaryotic release factor eRF1–eRF3-associated termination complex

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