Novel suppressors of alpha-synuclein toxicity identified using yeast.
The mechanism by which the Parkinson's disease-related protein alpha-synuclein (alpha-syn) causes neurodegeneration has not been elucidated. To determine the genes that protect cells from alpha-syn, we used a genetic screen to identify suppressors of the super sensitivity of the yeast Saccharomyces cerevisiae expressing alpha-syn to killing by hydrogen peroxide. Forty genes in ubiquitin-dependent protein catabolism, protein biosynthesis, vesicle trafficking and the response to stress were identified. Five of the forty genes--ENT3, IDP3, JEM1, ARG2 and HSP82--ranked highest in their ability to block alpha-syn-induced reactive oxygen species accumulation, and these five genes were characterized in more detail. The deletion of any of these five genes enhanced the toxicity of alpha-syn as judged by growth defects compared with wild-type cells expressing alpha-syn, which indicates that these genes protect cells from alpha-syn. Strikingly, four of the five genes are specific for alpha-syn in that they fail to protect cells from the toxicity of the two inherited mutants A30P or A53T. This finding suggests that alpha-syn causes toxicity to cells through a different pathway than these two inherited mutants. Lastly, overexpression of Ent3p, which is a clathrin adapter protein involved in protein transport between the Golgi and the vacuole, causes alpha-syn to redistribute from the plasma membrane into cytoplasmic vesicular structures. Our interpretation is that Ent3p mediates the transport of alpha-syn to the vacuole for proteolytic degradation. A similar clathrin adaptor protein, epsinR, exists in humans.
Liang
Jun
J
Department of Biochemistry and Molecular Biology, Louisiana State University Health Sciences Center, Shreveport, LA 71130-3932, USA.
Clark-Dixon
Cheryl
C
Wang
Shaoxiao
S
Flower
Todd R
TR
Williams-Hart
Tara
T
Zweig
Richard
R
Robinson
Lucy C
LC
Tatchell
Kelly
K
Witt
Stephan N
SN
D033942:Adaptor Proteins, Vesicular Transport / Q000235:genetics / Q000378:metabolism; D018841:HSP90 Heat-Shock Proteins / Q000235:genetics / Q000378:metabolism; D006801:Humans; D007521:Isocitrate Dehydrogenase / Q000235:genetics / Q000378:metabolism; D008565:Membrane Proteins / Q000235:genetics / Q000378:metabolism; D018832:Molecular Chaperones; D010300:Parkinson Disease / Q000235:genetics* / Q000378:metabolism; D021381:Protein Transport; D017382:Reactive Oxygen Species / Q000378:metabolism; D012441:Saccharomyces cerevisiae / Q000187:drug effects / Q000235:genetics* / Q000378:metabolism; D029701:Saccharomyces cerevisiae Proteins / Q000235:genetics / Q000378:metabolism; D013489:Suppression, Genetic; D051844:alpha-Synuclein / Q000235:genetics / Q000378:metabolism / Q000633:toxicity*
D033942:Adaptor Proteins, Vesicular Transport; C495587:Ent3 protein, S cerevisiae; C074510:HSP82 protein, S cerevisiae; D018841:HSP90 Heat-Shock Proteins; C106168:JEM1 protein, S cerevisiae; D008565:Membrane Proteins; D018832:Molecular Chaperones; D017382:Reactive Oxygen Species; D029701:Saccharomyces cerevisiae Proteins; D051844:alpha-Synuclein; D007521:Isocitrate Dehydrogenase; C039355:isocitrate dehydrogenase (NADP+)
Cell Mol Life Sci. 2000 Dec;57(13-14):1894-908
11215516
Biochim Biophys Acta. 2008 Oct;1783(10):1767-80
18634833
Proc Natl Acad Sci U S A. 2004 Jun 8;101(23):8658-63
15161972
Nat Neurosci. 2005 May;8(5):657-63
15834418
J Biol Chem. 1999 Oct 22;274(43):30527-33
10521434
Proc Natl Acad Sci U S A. 1997 Nov 25;94(24):12949-56
9371781
Proc Natl Acad Sci U S A. 2008 Jan 15;105(2):763-8
18178617
Am J Pathol. 1998 Apr;152(4):879-84
9546347
FASEB J. 2004 Apr;18(6):617-26
15054084
Biochemistry. 1998 Apr 7;37(14):4901-9
9538008
Biochemistry. 2001 Aug 21;40(33):9927-34
11502187
Nat Cell Biol. 2002 Feb;4(2):160-4
11813001
Science. 1997 Jun 27;276(5321):2045-7
9197268
Science. 2003 Dec 5;302(5651):1772-5
14657500
Cell Mol Life Sci. 2002 Oct;59(10):1640-8
12475174
Yeast. 1995 Jan;11(1):53-5
7762301
Science. 2003 Oct 31;302(5646):819-22
14593166
Mol Biol Cell. 1998 Nov;9(11):3071-83
9802897
Science. 2003 Oct 31;302(5646):841
14593171
Nat Med. 2002 Jun;8(6):600-6
12042811
Cell. 1980 Mar;19(3):753-64
6244896
Nat Genet. 1998 Feb;18(2):106-8
9462735
J Mol Biol. 2005 Sep 2;351(5):1081-100
16051265
Proc Natl Acad Sci U S A. 2008 Jan 15;105(2):728-33
18182484
Annu Rev Microbiol. 2005;59:407-50
16153175
J Cell Biol. 2007 Jun 18;177(6):1091-104
17576801
Annu Rev Neurosci. 2003;26:267-98
12704221
Mol Biol Cell. 2007 May;18(5):1803-15
17344475
J Biol Chem. 1997 May 16;272(20):12889-92
9148890
EMBO J. 1999 Aug 16;18(16):4383-93
10449404
Science. 2006 Jul 21;313(5785):324-8
16794039
PLoS Genet. 2008 Mar 21;4(3):e1000027
18369446
Hum Mol Genet. 2005 Dec 15;14(24):3801-11
16239241
Ann Neurol. 2004 Feb;55(2):164-73
14755719
FEMS Yeast Res. 2006 Dec;6(8):1107-16
17156009
Mol Biol Cell. 2003 Feb;14(2):625-41
12589059
J Bacteriol. 1978 Mar;133(3):1096-1107
205532
J Biol Chem. 2004 Mar 26;279(13):12924-34
14711827
J Biol Chem. 1998 Feb 6;273(6):3702-11
9452501
J Biol Chem. 1993 Jun 5;268(16):12116-22
8099357
Trends Genet. 2004 Jul;20(7):273-7
15219388
Mol Biol Cell. 2006 Sep;17(9):3907-20
16790491
Exp Neurol. 2005 Feb;191 Suppl 1:S17-27
15629758
PLoS Biol. 2004 Aug;2(8):E246
15314660
Nat Cell Biol. 2003 Jan;5(1):77-81
12483220
J Bacteriol. 1979 Dec;140(3):874-80
391804
Science. 2002 Feb 1;295(5556):865-8
11823645
Annu Rev Neurosci. 2005;28:57-87
16022590
Proc Natl Acad Sci U S A. 1997 May 13;94(10):4890-4
9144160
Science. 2003 Dec 5;302(5651):1769-72
14657499
EMBO Rep. 2004 Jun;5(6):567-71
15170475
Ann Bot. 2002 Jun;89 Spec No:825-32
12102508
Biochemistry. 2002 Apr 9;41(14):4595-602
11926821
Yeast. 1998 Jan 30;14(2):115-32
9483801
CRC Crit Rev Biochem. 1986;21(3):277-317
3536302
Cell. 2005 Mar 11;120(5):715-27
15766533
J Biol Chem. 2001 Nov 16;276(46):42869-80
11553611
Cell Tissue Res. 2004 Oct;318(1):185-8
15503154
Genetics. 2005 May;170(1):47-59
15744056
Mol Biol Cell. 2004 Jul;15(7):3031-41
15107463
J Cell Sci. 2004 Jan 1;117(Pt 1):9-18
14657269