Translation Initiation Factors eIF3 and HCR1 Control Translation Termination and Stop Codon Read-Through in Yeast Cells
Translation is divided into initiation, elongation, termination and ribosome recycling. Earlier work implicated several eukaryotic initiation factors (eIFs) in ribosomal recycling in vitro. Here, we uncover roles for HCR1 and eIF3 in translation termination in vivo. A substantial proportion of eIF3, HCR1 and eukaryotic release factor 3 (eRF3) but not eIF5 (a well-defined “initiation-specific” binding partner of eIF3) specifically co-sediments with 80S couples isolated from RNase-treated heavy polysomes in an eRF1-dependent manner, indicating the presence of eIF3 and HCR1 on terminating ribosomes. eIF3 and HCR1 also occur in ribosome- and RNA-free complexes with both eRFs and the recycling factor ABCE1/RLI1. Several eIF3 mutations reduce rates of stop codon read-through and genetically interact with mutant eRFs. In contrast, a slow growing deletion of hcr1 increases read-through and accumulates eRF3 in heavy polysomes in a manner suppressible by overexpressed ABCE1/RLI1. Based on these and other findings we propose that upon stop codon recognition, HCR1 promotes eRF3·GDP ejection from the post-termination complexes to allow binding of its interacting partner ABCE1/RLI1. Furthermore, the fact that high dosage of ABCE1/RLI1 fully suppresses the slow growth phenotype of hcr1Δ as well as its termination but not initiation defects implies that the termination function of HCR1 is more critical for optimal proliferation than its function in translation initiation. Based on these and other observations we suggest that the assignment of HCR1 as a bona fide eIF3 subunit should be reconsidered. Together our work characterizes novel roles of eIF3 and HCR1 in stop codon recognition, defining a communication bridge between the initiation and termination/recycling phases of translation.
Vyšlo v časopise:
Translation Initiation Factors eIF3 and HCR1 Control Translation Termination and Stop Codon Read-Through in Yeast Cells. PLoS Genet 9(11): e32767. doi:10.1371/journal.pgen.1003962
Kategorie:
Research Article
prolekare.web.journal.doi_sk:
https://doi.org/10.1371/journal.pgen.1003962
Souhrn
Translation is divided into initiation, elongation, termination and ribosome recycling. Earlier work implicated several eukaryotic initiation factors (eIFs) in ribosomal recycling in vitro. Here, we uncover roles for HCR1 and eIF3 in translation termination in vivo. A substantial proportion of eIF3, HCR1 and eukaryotic release factor 3 (eRF3) but not eIF5 (a well-defined “initiation-specific” binding partner of eIF3) specifically co-sediments with 80S couples isolated from RNase-treated heavy polysomes in an eRF1-dependent manner, indicating the presence of eIF3 and HCR1 on terminating ribosomes. eIF3 and HCR1 also occur in ribosome- and RNA-free complexes with both eRFs and the recycling factor ABCE1/RLI1. Several eIF3 mutations reduce rates of stop codon read-through and genetically interact with mutant eRFs. In contrast, a slow growing deletion of hcr1 increases read-through and accumulates eRF3 in heavy polysomes in a manner suppressible by overexpressed ABCE1/RLI1. Based on these and other findings we propose that upon stop codon recognition, HCR1 promotes eRF3·GDP ejection from the post-termination complexes to allow binding of its interacting partner ABCE1/RLI1. Furthermore, the fact that high dosage of ABCE1/RLI1 fully suppresses the slow growth phenotype of hcr1Δ as well as its termination but not initiation defects implies that the termination function of HCR1 is more critical for optimal proliferation than its function in translation initiation. Based on these and other observations we suggest that the assignment of HCR1 as a bona fide eIF3 subunit should be reconsidered. Together our work characterizes novel roles of eIF3 and HCR1 in stop codon recognition, defining a communication bridge between the initiation and termination/recycling phases of translation.
Zdroje
1. DongJ, LaiR, NielsenK, FeketeCA, QiuH, et al. (2004) The essential ATP-binding cassette protein RLI1 functions in translation by promoting preinitiation complex assembly. J Biol Chem 279: 42157–42168.
2. KhoshnevisS, GrossT, RotteC, BaierleinC, FicnerR, et al. (2010) The iron-sulphur protein RNase L inhibitor functions in translation termination. EMBO Rep 11: 214–219.
3. BolgerTA, FolkmannAW, TranEJ, WenteSR (2008) The mRNA export factor Gle1 and inositol hexakisphosphate regulate distinct stages of translation. Cell 134: 624–633.
4. SainiP, EylerDE, GreenR, DeverTE (2009) Hypusine-containing protein eIF5A promotes translation elongation. Nature 459: 118–121.
5. PisarevAV, HellenCUT, PestovaTV (2007) Recycling of Eukaryotic Posttermination Ribosomal Complexes. Cell 131: 286–299.
6. PisarevAV, SkabkinMA, PisarevaVP, SkabkinaOV, RakotondrafaraAM, et al. (2010) The Role of ABCE1 in Eukaryotic Posttermination Ribosomal Recycling. Mol Cell 37: 196–210.
7. ValášekLS (2012) ‘Ribozoomin’ – Translation Initiation from the Perspective of the Ribosome-bound Eukaryotic Initiation Factors (eIFs). Curr Protein Pept Sci 13: 305–330.
8. PöyryTA, KaminskiA, JacksonRJ (2004) What determines whether mammalian ribosomes resume scanning after translation of a short upstream open reading frame? Genes Dev 18: 62–75.
9. SzameczB, RutkaiE, CuchalovaL, MunzarovaV, HerrmannovaA, et al. (2008) eIF3a cooperates with sequences 5′ of uORF1 to promote resumption of scanning by post-termination ribosomes for reinitiation on GCN4 mRNA. Genes Dev 22: 2414–2425.
10. MunzarováV, PánekJ, GunišováS, DányiI, SzameczB, et al. (2011) Translation Reinitiation Relies on the Interaction between eIF3a/TIF32 and Progressively Folded cis-Acting mRNA Elements Preceding Short uORFs. PLoS Genet 7: e1002137.
11. AlkalaevaEZ, PisarevAV, FrolovaLY, KisselevLL, PestovaTV (2006) In vitro reconstitution of eukaryotic translation reveals cooperativity between release factors eRF1 and eRF3. Cell 125: 1125–1136.
12. ShoemakerCJ, GreenR (2011) Kinetic analysis reveals the ordered coupling of translation termination and ribosome recycling in yeast. Proc Natl Acad Sci U S A 108: E1392–1398.
13. BeckerT, FranckenbergS, WicklesS, ShoemakerCJ, AngerAM, et al. (2012) Structural basis of highly conserved ribosome recycling in eukaryotes and archaea. Nature 482: 501–506.
14. KeelingKM, LanierJ, DuM, Salas-MarcoJ, GaoL, et al. (2004) Leaky termination at premature stop codons antagonizes nonsense-mediated mRNA decay in S. cerevisiae. RNA 10: 691–703.
15. SchuldinerM, CollinsSR, ThompsonNJ, DenicV, BhamidipatiA, et al. (2005) Exploration of the function and organization of the yeast early secretory pathway through an epistatic miniarray profile. Cell 123: 507–519.
16. FirczukH, KannambathS, PahleJ, ClaydonA, BeynonR, et al. (2013) An in vivo control map for the eukaryotic mRNA translation machinery. Mol Syst Biol 9: 635.
17. BarthelmeD, DinkelakerS, AlbersSV, LondeiP, ErmlerU, et al. (2011) Ribosome recycling depends on a mechanistic link between the FeS cluster domain and a conformational switch of the twin-ATPase ABCE1. Proc Natl Acad Sci U S A 108: 3228–3233.
18. YaruninA, PanseVG, PetfalskiE, DezC, TollerveyD, et al. (2005) Functional link between ribosome formation and biogenesis of ironsulfur proteins. EMBO J 24: 580–588.
19. ValášekL, PhanL, SchoenfeldLW, ValáškováV, HinnebuschAG (2001) Related eIF3 subunits TIF32 and HCR1 interact with an RNA recoginition motif in PRT1 required for eIF3 integrity and ribosome binding. EMBO J 20: 891–904.
20. NielsenKH, ValášekL, SykesC, JivotovskayaA, HinnebuschAG (2006) Interaction of the RNP1 motif in PRT1 with HCR1 promotes 40S binding of eukaryotic initiation factor 3 in yeast. Mol Cell Biol 26: 2984–2998.
21. ElAntakL, WagnerS, HerrmannováA, KaráskováM, RutkaiE, et al. (2010) The indispensable N-terminal half of eIF3j co-operates with its structurally conserved binding partner eIF3b-RRM and eIF1A in stringent AUG selection. J Mol Biol 396: 1097–1116.
22. ChiuW-L, WagnerS, HerrmannováA, BurelaL, ZhangF, et al. (2010) The C-Terminal Region of Eukaryotic Translation Initiation Factor 3a (eIF3a) Promotes mRNA Recruitment, Scanning, and, Together with eIF3j and the eIF3b RNA Recognition Motif, Selection of AUG Start Codons. Mol Cell Biol 30: 4415–4434.
23. ValášekL, HašekJ, NielsenKH, HinnebuschAG (2001) Dual function of eIF3j/Hcr1p in processing 20 S Pre-rRNA and translation initiation. J Biol Chem 276: 43351–43360.
24. KovarikP, HašekJ, ValášekL, RuisH (1998) RPG1: an essential gene of saccharomyces cerevisiae encoding a 110-kDa protein required for passage through the G1 phase. Curr Genet 33: 100–109.
25. ValášekL, SzameczB, HinnebuschAG, NielsenKH (2007) In vivo stabilization of preinitiation complexes by formaldehyde cross-linking. Methods Enzymol 429: 163–183.
26. Burnicka-TurekO, KataA, BuyandelgerB, EbermannL, KramannN, et al. (2010) Pelota interacts with HAX1, EIF3G and SRPX and the resulting protein complexes are associated with the actin cytoskeleton. BMC Cell Biol 11: 28.
27. von der HaarT (2008) A quantitative estimation of the global translational activity in logarithmically growing yeast cells. BMC Syst Biol 2: 87.
28. Akhmaloka, SusilowatiPE, Subandi, MadayantiF (2008) Mutation at tyrosine in AMLRY (GILRY like) motif of yeast eRF1 on nonsense codons suppression and binding affinity to eRF3. Int J Biol Sci 4: 87–95.
29. BradleyME, BagriantsevS, VishveshwaraN, LiebmanSW (2003) Guanidine reduces stop codon read-through caused by missense mutations in SUP35 or SUP45. Yeast 20: 625–632.
30. BertramG, BellHA, RitchieDW, FullertonG, StansfieldI (2000) Terminating eukaryote translation: domain 1 of release factor eRF1 functions in stop codon recognition. Rna 6: 1236–1247.
31. MerrittGH, NaemiWR, MugnierP, WebbHM, TuiteMF, et al. (2010) Decoding accuracy in eRF1 mutants and its correlation with pleiotropic quantitative traits in yeast. Nucleic Acids Res 38: 5479–5492.
32. Harel-SharvitL, EldadN, HaimovichG, BarkaiO, DuekL, et al. (2010) RNA polymerase II subunits link transcription and mRNA decay to translation. Cell 143: 552–563.
33. IskenO, KimYK, HosodaN, MayeurGL, HersheyJWB, et al. (2008) Upf1 Phosphorylation Triggers Translational Repression during Nonsense-Mediated mRNA Decay. Cell 133: 314–327.
34. ShaZ, BrillLM, CabreraR, KleifeldO, ScheligaJS, et al. (2009) The eIF3 interactome reveals the translasome, a supercomplex linking protein synthesis and degradation machineries. Mol Cell 36: 141–152.
35. Querol-AudiJ, SunC, VoganJM, SmithMD, GuY, et al. (2013) Architecture of human translation initiation factor 3. Structure 21: 920–928.
36. HashemY, des GeorgesA, DhoteV, LangloisR, LiaoHY, et al. (2013) Structure of the Mammalian Ribosomal 43S Preinitiation Complex Bound to the Scanning Factor DHX29. Cell 153: 1108–1119.
37. ZhouM, SandercockAM, FraserCS, RidlovaG, StephensE, et al. (2008) Mass spectrometry reveals modularity and a complete subunit interaction map of the eukaryotic translation factor eIF3. Proc Natl Acad Sci USA 105: 18139–18144.
38. FraserCS, BerryKE, HersheyJW, DoudnaJA (2007) 3j is located in the decoding center of the human 40S ribosomal subunit. Mol Cell 26: 811–819.
39. PisarevAV, KolupaevaVG, YusupovMM, HellenCUT, PestovaTV (2008) Ribosomal position and contacts of mRNA in eukaryotic translation initiation complexes. EMBO J 27: 1609–1621.
40. MasutaniM, SonenbergN, YokoyamaS, ImatakaH (2007) Reconstitution reveals the functional core of mammalian eIF3. EMBO J 26: 3373–3383.
41. TaylorD, UnbehaunA, LiW, DasS, LeiJ, et al. (2012) Cryo-EM structure of the mammalian eukaryotic release factor eRF1-eRF3-associated termination complex. Proc Natl Acad Sci U S A 109: 18413–18418.
42. CuchalováL, KoubaT, HerrmannováA, DanyiI, ChiuW-l, et al. (2010) The RNA Recognition Motif of Eukaryotic Translation Initiation Factor 3g (eIF3g) Is Required for Resumption of Scanning of Posttermination Ribosomes for Reinitiation on GCN4 and Together with eIF3i Stimulates Linear Scanning. Mol Cell Biol 30: 4671–4686.
43. HerrmannováA, DaujotyteD, YangJC, CuchalováL, GorrecF, et al. (2012) Structural analysis of an eIF3 subcomplex reveals conserved interactions required for a stable and proper translation pre-Initiation complex assembly. Nucleic Acids Res 40: 2294–2311.
44. ValášekL, MathewA, ShinBS, NielsenKH, SzameczB, et al. (2003) The Yeast eIF3 Subunits TIF32/a and NIP1/c and eIF5 Make Critical Connections with the 40S Ribosome in vivo. Genes Dev 17: 786–799.
45. KongC, ItoK, WalshMA, WadaM, LiuY, et al. (2004) Crystal structure and functional analysis of the eukaryotic class II release factor eRF3 from S. pombe. Mol Cell 14: 233–245.
46. ValášekL, HašekJ, TrachselH, ImreEM, RuisH (1999) The Saccharomyces cerevisiae HCRI gene encoding a homologue of the p35 subunit of human translation eukaryotic initiation factor 3 (eIF3) is a high copy suppressor of a temperature-sensitive mutation in the Rpg1p subunit of yeast eIF3. J Biol Chem 274: 27567–27572.
47. TarunSZ, SachsAB (1996) Association of the yeast poly(A) tail binding protein with translation initiation factor eIF-4G. EMBO J 15: 7168–7177.
48. MuhlradD, ParkerR (1999) Recognition of yeast mRNAs as “nonsense containing” leads to both inhibition of mRNA translation and mRNA degradation: implications for the control of mRNA decapping. Mol Biol Cell 10: 3971–3978.
49. NielsenKH, ValášekL (2007) In vivo deletion analysis of the architecture of a multi-protein complex of translation initiation factors. Methods Enzymol 431: 15–32.
50. ValášekL, TrachselH, HašekJ, RuisH (1998) Rpg1, the Saccharomyces cerevisiae homologue of the largest subunit of mammlian translation initiation factor 3, is required for translational activity. J Biol Chem 273: 21253–21260.
51. GrantCM, HinnebuschAG (1994) Effect of sequence context at stop codons on efficiency of reinitiation in GCN4 translational control. Mol Cell Biol 14: 606–618.
Štítky
Genetika Reprodukčná medicínaČlánok vyšiel v časopise
PLOS Genetics
2013 Číslo 11
- Gynekologové a odborníci na reprodukční medicínu se sejdou na prvním virtuálním summitu
- Je „freeze-all“ pro všechny? Odborníci na fertilitu diskutovali na virtuálním summitu
Najčítanejšie v tomto čísle
- Genetic and Functional Studies Implicate Synaptic Overgrowth and Ring Gland cAMP/PKA Signaling Defects in the Neurofibromatosis-1 Growth Deficiency
- RNA∶DNA Hybrids Initiate Quasi-Palindrome-Associated Mutations in Highly Transcribed Yeast DNA
- The Light Skin Allele of in South Asians and Europeans Shares Identity by Descent
- Roles of XRCC2, RAD51B and RAD51D in RAD51-Independent SSA Recombination