title: Fusel alcohols regulate translation initiation by inhibiting eIF2B to reduce ternary complex in a mechanism that may involve altering the integrity and dynamics of the eIF2B body.
abstract: Recycling of eIF2-GDP to the GTP-bound form constitutes a core essential, regulated step in eukaryotic translation. This reaction is mediated by eIF2B, a heteropentameric factor with important links to human disease. eIF2 in the GTP-bound form binds to methionyl initiator tRNA to form a ternary complex, and the levels of this ternary complex can be a critical determinant of the rate of protein synthesis. Here we show that eIF2B serves as the target for translation inhibition by various fusel alcohols in yeast. Fusel alcohols are endpoint metabolites from amino acid catabolism, which signal nitrogen scarcity. We show that the inhibition of eIF2B leads to reduced ternary complex levels and that different eIF2B subunit mutants alter fusel alcohol sensitivity. A DNA tiling array strategy was developed that overcame difficulties in the identification of these mutants where the phenotypic distinctions were too subtle for classical complementation cloning. Fusel alcohols also lead to eIF2alpha dephosphorylation in a Sit4p-dependent manner. In yeast, eIF2B occupies a large cytoplasmic body where guanine nucleotide exchange on eIF2 can occur and be regulated. Fusel alcohols impact on both the movement and dynamics of this 2B body. Overall, these results confirm that the guanine nucleotide exchange factor, eIF2B, is targeted by fusel alcohols. Moreover, they highlight a potential connection between the movement or integrity of the 2B body and eIF2B regulation.
authors.affiliation: Faculty of Life Sciences, University of Manchester, Manchester M13 9PT, United Kingdom.
authors.forename: Eleanor J
authors.initials: EJ
authors.lastname: Taylor
authors.forename: Susan G
authors.initials: SG
authors.lastname: Campbell
authors.forename: Christian D
authors.initials: CD
authors.lastname: Griffiths
authors.forename: Peter J
authors.initials: PJ
authors.lastname: Reid
authors.forename: John W
authors.initials: JW
authors.lastname: Slaven
authors.forename: Richard J
authors.initials: RJ
authors.lastname: Harrison
authors.forename: Paul F G
authors.initials: PF
authors.lastname: Sims
authors.forename: Graham D
authors.initials: GD
authors.lastname: Pavitt
authors.forename: Daniela
authors.initials: D
authors.lastname: Delneri
authors.forename: Mark P
authors.initials: MP
authors.lastname: Ashe
mesh_terms: D000438:Alcohols / Q000378:metabolism / Q000494:pharmacology*; D001483:Base Sequence; D020717:Eukaryotic Initiation Factor-2B / Q000037:antagonists & inhibitors* / Q000737:chemistry / Q000235:genetics / Q000378:metabolism*; D006153:Guanosine Diphosphate / Q000378:metabolism; D006160:Guanosine Triphosphate / Q000378:metabolism; D006801:Humans; D008969:Molecular Sequence Data; D046912:Multiprotein Complexes; D009154:Mutation; D014176:Protein Biosynthesis / Q000187:drug effects*; D054648:Protein Phosphatase 2 / Q000235:genetics / Q000378:metabolism; D021122:Protein Subunits / Q000737:chemistry / Q000235:genetics / Q000378:metabolism*; D011993:Recombinant Fusion Proteins / Q000235:genetics / Q000378:metabolism; D012441:Saccharomyces cerevisiae / Q000235:genetics / Q000378:metabolism; D029701:Saccharomyces cerevisiae Proteins / Q000737:chemistry / Q000235:genetics / Q000378:metabolism
chemical_list: D000438:Alcohols; D020717:Eukaryotic Initiation Factor-2B; D046912:Multiprotein Complexes; D021122:Protein Subunits; D011993:Recombinant Fusion Proteins; D029701:Saccharomyces cerevisiae Proteins; D006153:Guanosine Diphosphate; C007807:fusel oil; D006160:Guanosine Triphosphate; D054648:Protein Phosphatase 2; C104343:SIT4 protein, S cerevisiae
references.citation: Mol Cell Biol. 1993 Mar;13(3):1920-32
references.pmid: 8441423
references.citation: Mol Cell Biol. 1996 Nov;16(11):6603-16
references.pmid: 8887689
references.citation: J Biol Chem. 1988 Apr 25;263(12):5526-33
references.pmid: 3356695
references.citation: J Biol Chem. 2002 Jun 14;277(24):21542-8
references.pmid: 11940596
references.citation: Proc Natl Acad Sci U S A. 2004 Aug 3;101(31):11269-74
references.pmid: 15277680
references.citation: Alcohol Clin Exp Res. 1999 Sep;23(9):1552-60
references.pmid: 10512323
references.citation: FEMS Yeast Res. 2004 Oct;5(1):43-9
references.pmid: 15381121
references.citation: Genes Dev. 1996 Aug 1;10(15):1904-16
references.pmid: 8756348
references.citation: J Cell Biol. 2001 May 28;153(5):1011-22
references.pmid: 11381086
references.citation: Mol Cell Biol. 1988 Nov;8(11):4808-20
references.pmid: 3062370
references.citation: J Cell Sci. 2003 Aug 15;116(Pt 16):3423-31
references.pmid: 12840070
references.citation: Mol Cell Biol. 2008 Mar;28(5):1429-42
references.pmid: 18160716
references.citation: Acta Crystallogr D Biol Crystallogr. 2004 Sep;60(Pt 9):1600-5
references.pmid: 15333931
references.citation: Genes Dev. 1994 May 1;8(9):1087-105
references.pmid: 7926789
references.citation: Mol Cell Biol. 1993 Aug;13(8):4618-31
references.pmid: 8336705
references.citation: Curr Opin Cell Biol. 2005 Apr;17(2):197-202
references.pmid: 15780597
references.citation: Genes Dev. 2003 Apr 1;17(7):859-72
references.pmid: 12654728
references.citation: Mol Biol Cell. 2000 Mar;11(3):833-48
references.pmid: 10712503
references.citation: Mol Biol Cell. 2005 Aug;16(8):3727-39
references.pmid: 15930127
references.citation: Mol Cell Biol. 1998 Apr;18(4):1967-77
references.pmid: 9528768
references.citation: Arch Biochem Biophys. 2005 Jan 1;433(1):85-95
references.pmid: 15581568
references.citation: Am J Physiol Endocrinol Metab. 2006 Jun;290(6):E1267-75
references.pmid: 16434554
references.citation: Biochem Soc Trans. 2005 Dec;33(Pt 6):1487-92
references.pmid: 16246152
references.citation: Yeast. 2004 Aug;21(11):947-62
references.pmid: 15334558
references.citation: Biochem Soc Trans. 2006 Feb;34(Pt 1):7-11
references.pmid: 16246168
references.citation: J Cell Biol. 2005 Sep 12;170(6):925-34
references.pmid: 16157703
references.citation: Cell Cycle. 2006 Apr;5(7):678-80
references.pmid: 16582624
references.citation: EMBO J. 2001 Nov 15;20(22):6464-74
references.pmid: 11707417
references.citation: Mol Cell. 2003 Mar;11(3):619-33
references.pmid: 12667446
references.citation: Annu Rev Microbiol. 2005;59:407-50
references.pmid: 16153175
references.citation: J Cell Biol. 2004 Oct 11;167(1):27-33
references.pmid: 15479734
references.citation: Protein Sci. 2006 Sep;15(9):2107-19
references.pmid: 16943444
references.citation: Mol Cell Biol. 1992 Dec;12(12):5700-10
references.pmid: 1333044
references.citation: J Cell Biol. 2003 Nov 24;163(4):767-75
references.pmid: 14638860
references.citation: Mol Cell Biol. 2001 Aug;21(15):5018-30
references.pmid: 11438658
references.citation: Genes Dev. 1999 Nov 15;13(22):2946-57
references.pmid: 10580002
references.citation: Biochem J. 2001 May 1;355(Pt 3):609-15
references.pmid: 11311121
references.citation: Cell. 1992 Feb 7;68(3):585-96
references.pmid: 1739968
references.citation: Genetics. 2001 Jun;158(2):549-62
references.pmid: 11404321
references.citation: Semin Cell Dev Biol. 2005 Feb;16(1):3-12
references.pmid: 15659334
references.citation: Eukaryot Cell. 2002 Dec;1(6):1032-40
references.pmid: 12477803
references.citation: Genome Res. 2000 Aug;10(8):1172-84
references.pmid: 10958635
references.citation: Mol Biol Cell. 2001 Dec;12(12):4103-13
references.pmid: 11739804
references.citation: Mol Cell Biol. 2007 May;27(9):3378-89
references.pmid: 17325030
references.citation: Nat Cell Biol. 2002 Jul;4(7):502-8
references.pmid: 12068306
references.citation: Genes Dev. 1998 Feb 15;12(4):514-26
references.pmid: 9472020
references.citation: Appl Environ Microbiol. 2008 Apr;74(8):2259-66
references.pmid: 18281432
references.citation: J Mol Biol. 2007 Jul 6;370(2):315-30
references.pmid: 17512538
references.citation: Methods Enzymol. 1991;194:195-230
references.pmid: 2005788
references.citation: Mol Cell Biol. 2004 Apr;24(8):3295-306
references.pmid: 15060152
references.citation: Neurol Sci. 2006 Sep;27(4):271-7
references.pmid: 16998732
references.citation: Mol Cell Biol. 2004 Oct;24(19):8332-41
references.pmid: 15367655
references.citation: Mol Biol Cell. 2000 Jan;11(1):183-99
references.pmid: 10637301
references.citation: Science. 2006 Mar 31;311(5769):1932-6
references.pmid: 16527929
references.citation: Mol Cell Biol. 2005 Nov;25(21):9340-9
references.pmid: 16227585
references.citation: Mol Cell Biol. 1997 Mar;17(3):1298-313
references.pmid: 9032257
references.citation: Mol Cell Biol. 2007 Jul;27(14):5225-34
references.pmid: 17526738
references.citation: Mol Cell Biol. 2004 Mar;24(6):2352-63
references.pmid: 14993275