title: Assessing the subcellular dynamics of alpha-synuclein using photoactivation microscopy.
abstract: Alpha-synuclein (aSyn) is implicated in Parkinson's disease and several other neurodegenerative disorders. To date, the function and intracellular dynamics of aSyn are still unclear. Here, we tracked the dynamics of aSyn using photoactivatable green fluorescent protein as a reporter. We found that the availability of the aSyn N terminus modulates its shuttling into the nucleus. Interestingly, familial aSyn mutations altered the dynamics at which the protein distributes throughout the cell. Both the A30P and A53T aSyn mutations increase the speed at which the protein moves between the nucleus and cytoplasm, respectively. We also found that specific kinases potentiate the shuttling of aSyn between nucleus and cytoplasm. A mutant aSyn form that blocks S129 phosphorylation, S129A, results in the formation of cytoplasmic inclusions, suggesting phosphorylation modulates aggregation in addition to modulating aSyn intracellular dynamics. Finally, we found that the molecular chaperone HSP70 accelerates the entry of aSyn into the nuclear compartment.
authors: Cell and Molecular Neuroscience Unit, Instituto de Medicina Molecular, Av. Prof. Egas Moniz, 1649-028 Lisboa, Portugal.
authors: Susana
authors: S
authors: Gonçalves
authors: Tiago Fleming
authors: TF
authors: Outeiro
mesh_terms: D002467:Cell Nucleus / Q000378:metabolism; D002470:Cell Survival; D054774:G-Protein-Coupled Receptor Kinase 5 / Q000378:metabolism; D049452:Green Fluorescent Proteins / Q000378:metabolism; D018840:HSP70 Heat-Shock Proteins / Q000378:metabolism; D006801:Humans; D008027:Light; D008853:Microscopy / Q000379:methods*; D009154:Mutation / Q000235:genetics; D010300:Parkinson Disease / Q000235:genetics / Q000473:pathology; D010766:Phosphorylation; D010768:Phosphoserine / Q000378:metabolism; D021381:Protein Transport; D013347:Subcellular Fractions / Q000378:metabolism; D051844:alpha-Synuclein / Q000737:chemistry / Q000378:metabolism*
chemical_list: D018840:HSP70 Heat-Shock Proteins; D051844:alpha-Synuclein; D049452:Green Fluorescent Proteins; D010768:Phosphoserine; D054774:G-Protein-Coupled Receptor Kinase 5
references: J Mol Biol. 2009 Jun 5;389(2):413-24
references: 19285989
references: Nat Genet. 1998 Feb;18(2):106-8
references: 9462735
references: Science. 2002 Sep 13;297(5588):1873-7
references: 12228718
references: J Biol Chem. 2000 Jan 7;275(1):390-7
references: 10617630
references: Proc Natl Acad Sci U S A. 2004 Nov 23;101(47):16466-71
references: 15536128
references: Ann Neurol. 2004 Feb;55(2):164-73
references: 14755719
references: J Biol Chem. 2000 Aug 25;275(34):26515-22
references: 10852916
references: J Biol Chem. 2009 Jan 30;284(5):2598-2602
references: 19004816
references: Nature. 2011 Aug 14;477(7362):107-10
references: 21841800
references: Biochemistry. 1996 Oct 29;35(43):13709-15
references: 8901511
references: Mol Neurodegener. 2010 Feb 09;5(1):9
references: 20181133
references: J Biol Chem. 2006 Oct 6;281(40):29739-52
references: 16847063
references: Science. 1997 Jun 27;276(5321):2045-7
references: 9197268
references: Nat Cell Biol. 2002 Feb;4(2):160-4
references: 11813001
references: Parkinsons Dis. 2011;2011:392180
references: 21826257
references: Free Radic Biol Med. 2012 Aug 15;53(4):993-1003
references: 22705949
references: Mol Cell Neurosci. 2005 Feb;28(2):326-34
references: 15691713
references: J Biol Chem. 2003 Apr 25;278(17):15313-8
references: 12586824
references: Biochemistry. 2010 Feb 9;49(5):862-71
references: 20041693
references: Science. 2003 Oct 31;302(5646):841
references: 14593171
references: J Biol Chem. 2005 Apr 15;280(15):14733-40
references: 15671022
references: J Biol Chem. 2005 Dec 30;280(52):43179-87
references: 16166095
references: PLoS One. 2008 Apr 02;3(4):e1867
references: 18382657
references: J Biol Chem. 2000 Nov 3;275(44):34393-8
references: 10952980
references: Hum Mol Genet. 2006 Oct 15;15(20):3012-23
references: 16959795
references: J Biol Chem. 2003 May 9;278(19):16873-7
references: 12621030
references: Hum Mol Genet. 2007 Mar 15;16(6):651-66
references: 17309880
references: Nat Neurosci. 2005 May;8(5):657-63
references: 15834418
references: J Neurosci. 2012 Feb 1;32(5):1536-44
references: 22302797
references: Biochemistry. 2003 Jul 22;42(28):8530-40
references: 12859200
references: J Biol Chem. 2005 Mar 11;280(10):9595-603
references: 15615727
references: Nat Protoc. 2011 Dec 15;7(1):62-8
references: 22179592
references: J Mol Biol. 2005 Sep 2;351(5):1081-100
references: 16051265
references: Mol Biol Cell. 2009 Aug;20(16):3725-39
references: 19553474
references: Nature. 1997 Aug 28;388(6645):839-40
references: 9278044
references: J Neurosci. 2004 Jul 28;24(30):6715-23
references: 15282274
references: PLoS One. 2010 May 11;5(5):e10589
references: 20485674
references: J Am Chem Soc. 2012 Mar 21;134(11):5196-210
references: 22339654
references: Biochemistry. 2003 Jul 22;42(28):8465-71
references: 12859192
references: Nat Cell Biol. 2003 Sep;Suppl:S7-14
references: 14562845
references: J Chem Neuroanat. 2011 Mar;41(2):97-110
references: 21172422
references: J Biol Chem. 2010 Jun 4;285(23):17648-61
references: 20356833
references: Science. 2002 Feb 1;295(5556):865-8
references: 11823645
references: J Biol Chem. 2004 Sep 10;279(37):39155-64
references: 15234983
references: FASEB J. 2006 Oct;20(12):2050-7
references: 17012257
references: PLoS One. 2009 Sep 04;4(9):e6906
references: 19730729
references: J Mol Biol. 2002 Oct 4;322(5):1089-102
references: 12367530
references: J Biol Chem. 2004 Jun 11;279(24):25497-502
references: 15044495
references: Neuroscience. 2001;104(3):901-12
references: 11440819
references: Proc Natl Acad Sci U S A. 2011 Oct 25;108(43):17797-802
references: 22006323
references: J Biol Chem. 2010 Jan 22;285(4):2807-22
references: 19889641
references: Biopolymers. 2010 Mar;93(3):218-28
references: 19768775
references: J Neurosci. 2008 Apr 16;28(16):4250-60
references: 18417705