title: Insight into molecular basis of curing of [PSI+] prion by overexpression of 104-kDa heat shock protein (Hsp104).
abstract: Yeast prions are a powerful model for understanding the dynamics of protein aggregation associated with a number of human neurodegenerative disorders. The AAA+ protein disaggregase Hsp104 can sever the amyloid fibrils produced by yeast prions. This action results in the propagation of "seeds" that are transmitted to daughter cells during budding. Overexpression of Hsp104 eliminates the [PSI+] prion but not other prions. Using biochemical methods we identified Hsp104 binding sites in the highly charged middle domain of Sup35, the protein determinant of [PSI+]. Deletion of a short segment of the middle domain (amino acids 129-148) diminishes Hsp104 binding and strongly affects the ability of the middle domain to stimulate the ATPase activity of Hsp104. In yeast, [PSI+] maintained by Sup35 lacking this segment, like other prions, is propagated by Hsp104 but cannot be cured by Hsp104 overexpression. These results provide new insight into the enigmatic specificity of Hsp104-mediated curing of yeast prions and sheds light on the limitations of the ability of Hsp104 to eliminate aggregates produced by other aggregation-prone proteins.
authors.affiliation: Department of Biochemistry, University of Toronto, Toronto, Ontario M5S 1A8, Canada.
authors.forename: Christopher W
authors.initials: CW
authors.lastname: Helsen
authors.affiliation: Department of Biochemistry, University of Toronto, Toronto, Ontario M5S 1A8, Canada. Electronic address: john.glover@utoronto.ca.
authors.forename: John R
authors.initials: JR
authors.lastname: Glover
mesh_terms: D000251:Adenosine Triphosphatases / Q000037:antagonists & inhibitors / Q000378:metabolism*; D000595:Amino Acid Sequence; D015966:Gene Expression Regulation, Fungal; D019791:Guanidine / Q000494:pharmacology; D006360:Heat-Shock Proteins / Q000037:antagonists & inhibitors / Q000378:metabolism*; D006801:Humans; D008969:Molecular Sequence Data; D009687:Nuclear Proteins / Q000737:chemistry / Q000378:metabolism*; D010446:Peptide Fragments / Q000737:chemistry / Q000378:metabolism; D011328:Prions / Q000737:chemistry / Q000378:metabolism*; D017434:Protein Structure, Tertiary; D021381:Protein Transport; D012568:Schizosaccharomyces / Q000235:genetics / Q000378:metabolism*; D029702:Schizosaccharomyces pombe Proteins / Q000737:chemistry / Q000378:metabolism*
chemical_list: D006360:Heat-Shock Proteins; D009687:Nuclear Proteins; D010446:Peptide Fragments; D011328:Prions; C093988:Psi protein, S pombe; D029702:Schizosaccharomyces pombe Proteins; D000251:Adenosine Triphosphatases; C546584:HSP104 protein, S pombe; D019791:Guanidine
references.citation: J Mol Biol. 2010 Sep 10;402(1):30-7
references.pmid: 20654624
references.citation: Yeast. 2001 Apr;18(6):489-97
references.pmid: 11284005
references.citation: Cell. 1998 Jul 10;94(1):73-82
references.pmid: 9674429
references.citation: PLoS One. 2009;4(3):e4670
references.pmid: 19262693
references.citation: PLoS Biol. 2004 Apr;2(4):E86
references.pmid: 15045026
references.citation: J Mol Biol. 2007 Jul 20;370(4):674-86
references.pmid: 17543332
references.citation: Cell. 2007 Dec 28;131(7):1366-77
references.pmid: 18160044
references.citation: Genetics. 2005 Mar;169(3):1227-42
references.pmid: 15545639
references.citation: Biochemistry. 2004 Jun 29;43(25):8107-15
references.pmid: 15209506
references.citation: Nature. 2004 Sep 9;431(7005):184-7
references.pmid: 15311209
references.citation: PLoS One. 2009 Sep 11;4(9):e6939
references.pmid: 19759825
references.citation: Biochem Biophys Res Commun. 1967 May 5;27(3):299-304
references.pmid: 6035110
references.citation: Proc Natl Acad Sci U S A. 2011 Mar 29;108(13):5337-41
references.pmid: 21402947
references.citation: Curr Microbiol. 2001 Jul;43(1):7-10
references.pmid: 11375656
references.citation: Genetics. 1996 Dec;144(4):1375-86
references.pmid: 8978027
references.citation: Eukaryot Cell. 2006 Feb;5(2):217-25
references.pmid: 16467463
references.citation: J Biol Chem. 2004 Feb 27;279(9):7378-83
references.pmid: 14668331
references.citation: J Biol Chem. 2008 Oct 31;283(44):30139-50
references.pmid: 18755692
references.citation: J Biol Chem. 2003 Dec 5;278(49):49636-43
references.pmid: 14507919
references.citation: Proc Natl Acad Sci U S A. 2011 Apr 26;108(17):6915-20
references.pmid: 21474779
references.citation: Proc Natl Acad Sci U S A. 2010 May 4;107(18):8135-40
references.pmid: 20404203
references.citation: J Mol Biol. 2011 May 6;408(3):432-48
references.pmid: 21392508
references.citation: Biochem Soc Symp. 2001;(68):35-43
references.pmid: 11573346
references.citation: Nucleic Acids Res. 1994 Dec 25;22(25):5767-8
references.pmid: 7838736
references.citation: PLoS One. 2011;6(7):e21953
references.pmid: 21760933
references.citation: Biochem Cell Biol. 2010 Feb;88(1):1-13
references.pmid: 20130674
references.citation: Yeast. 1995 Apr 15;11(4):355-60
references.pmid: 7785336
references.citation: Nature. 2005 Jun 9;435(7043):765-72
references.pmid: 15944694
references.citation: EMBO J. 1995 Sep 1;14(17):4365-73
references.pmid: 7556078
references.citation: EMBO J. 2001 Dec 3;20(23):6683-91
references.pmid: 11726504
references.citation: Genetics. 1997 Oct;147(2):507-19
references.pmid: 9335589
references.citation: Proc Natl Acad Sci U S A. 2005 Sep 6;102(36):12789-94
references.pmid: 16123122
references.citation: J Biol Chem. 2004 Aug 20;279(34):35692-701
references.pmid: 15178690
references.citation: Biochemistry. 2008 Feb 19;47(7):1918-27
references.pmid: 18197703
references.citation: Genes Cells. 2003 Dec;8(12):925-39
references.pmid: 14750948
references.citation: Cell. 2004 Nov 24;119(5):653-65
references.pmid: 15550247
references.citation: Nat Cell Biol. 2009 Mar;11(3):344-9
references.pmid: 19219034
references.citation: Genetics. 2001 Oct;159(2):527-35
references.pmid: 11606530
references.citation: Nature. 2000 Sep 28;407(6803):477-83
references.pmid: 11028992
references.citation: Mol Cell Biol. 2001 Jul;21(14):4656-69
references.pmid: 11416143
references.citation: J Biol Chem. 1998 Jun 19;273(25):15546-52
references.pmid: 9624144
references.citation: Genetics. 2006 Feb;172(2):827-35
references.pmid: 16272413
references.citation: Nature. 2004 Mar 18;428(6980):323-8
references.pmid: 15029196
references.citation: EMBO J. 2008 Oct 22;27(20):2712-24
references.pmid: 18833196
references.citation: Eukaryot Cell. 2005 Feb;4(2):289-97
references.pmid: 15701791
references.citation: Curr Biol. 2000 Nov 16;10(22):1443-6
references.pmid: 11102806
references.citation: Science. 2004 Jun 18;304(5678):1793-7
references.pmid: 15155912
references.citation: Mol Cell. 2006 Aug 4;23(3):425-38
references.pmid: 16885031
references.citation: Genes Dev. 1992 Jul;6(7):1153-64
references.pmid: 1628823
references.citation: Nature. 2007 Sep 13;449(7159):233-7
references.pmid: 17767153
references.citation: Curr Genet. 2006 Jan;49(1):21-9
references.pmid: 16307272
references.citation: Mol Cell. 2002 Apr;9(4):751-60
references.pmid: 11983167
references.citation: Methods Enzymol. 2010;470:709-34
references.pmid: 20946833
references.citation: J Biol Chem. 2004 Jul 9;279(28):29139-46
references.pmid: 15128736
references.citation: Nat Genet. 2008 Apr;40(4):460-5
references.pmid: 18362884
references.citation: EMBO J. 2000 Feb 1;19(3):324-31
references.pmid: 10654931
references.citation: Proc Natl Acad Sci U S A. 1999 Feb 16;96(4):1498-503
references.pmid: 9990052
references.citation: EMBO J. 2001 Nov 15;20(22):6236-45
references.pmid: 11707395
references.citation: EMBO J. 2001 May 1;20(9):2111-9
references.pmid: 11331577
references.citation: Biochem Cell Biol. 2010 Feb;88(1):63-75
references.pmid: 20130680
references.citation: Nature. 2008 Jan 17;451(7176):363-7
references.pmid: 18202664
references.citation: Nature. 1994 Dec 1;372(6505):475-8
references.pmid: 7984243
references.citation: Mol Cell Biol. 1999 Feb;19(2):1325-33
references.pmid: 9891066
references.citation: Science. 2010 Oct 29;330(6004):680-3
references.pmid: 21030659
references.citation: Science. 1996 Aug 2;273(5275):622-6
references.pmid: 8662547
references.citation: Cell. 1997 May 30;89(5):811-9
references.pmid: 9182769
references.citation: Science. 1995 May 12;268(5212):880-4
references.pmid: 7754373
references.citation: Science. 2000 Aug 25;289(5483):1317-21
references.pmid: 10958771
references.citation: Biochem Biophys Res Commun. 2010 Jan 1;391(1):1056-61
references.pmid: 19995551
references.citation: EMBO J. 2006 Feb 22;25(4):822-33
references.pmid: 16467849
references.citation: Proc Natl Acad Sci U S A. 2002 Dec 10;99 Suppl 4:16446-53
references.pmid: 12461168