S-Nitrosylation of DRP1 does not affect enzymatic activity and is not specific to Alzheimer's disease.
Mitochondrial dysfunction and synaptic loss are among the earliest events linked to Alzheimer's disease (AD) and might play a causative role in disease onset and progression. The underlying mechanisms of mitochondrial and synaptic dysfunction in AD remain unclear. We previously reported that nitric oxide (NO) triggers persistent mitochondrial fission and causes neuronal cell death. A recent article claimed that S-nitrosylation of dynamin related protein 1 (DRP1) at cysteine 644 causes protein dimerization and increased GTPase activity and is the mechanism responsible for NO-induced mitochondrial fission and neuronal injury in AD, but not in Parkinson's disease (PD). However, this report remains controversial. To resolve the controversy, we investigated the effects of S-nitrosylation on DRP1 structure and function. Contrary to the previous report, S-nitrosylation of DRP1 does not increase GTPase activity or cause dimerization. In fact, DRP1 does not exist as a dimer under native conditions, but rather as a tetramer capable of self-assembly into higher order spiral- and ring-like oligomeric structures after nucleotide binding. S-nitrosylation, as confirmed by the biotin-switch assay, has no impact on DRP1 oligomerization. Importantly, we found no significant difference in S-nitrosylated DRP1 (SNO-DRP1) levels in brains of age-matched normal, AD, or PD patients. We also found that S-nitrosylation is not specific to DRP1 because S-nitrosylated optic atrophy 1 (SNO-OPA1) is present at comparable levels in all human brain samples. Finally, we show that NO triggers DRP1 phosphorylation at serine 616, which results in its activation and recruitment to mitochondria. Our data indicate the mechanism underlying nitrosative stress-induced mitochondrial fragmentation in AD is not DRP1 S-nitrosylation.
Bossy
Blaise
B
Burnett School of Biomedical Sciences, College of Medicine, University of Central Florida, Orlando, FL, USA.
Petrilli
Alejandra
A
Klinglmayr
Eva
E
Chen
Jin
J
Lütz-Meindl
Ursula
U
Knott
Andrew B
AB
Masliah
Eliezer
E
Schwarzenbacher
Robert
R
Bossy-Wetzel
Ella
E
D000375:Aging / Q000473:pathology; D000544:Alzheimer Disease / Q000201:enzymology* / Q000209:etiology / Q000235:genetics / Q000473:pathology; D001710:Biotin / Q000378:metabolism; D001921:Brain / Q000201:enzymology* / Q000648:ultrastructure; D002461:Cell Line, Transformed; D003545:Cysteine / Q000235:genetics / Q000378:metabolism; D004229:Dithiothreitol / Q000494:pharmacology; D034281:Dynamins; D020558:GTP Phosphohydrolases / Q000235:genetics / Q000378:metabolism*; D006801:Humans; D046529:Microscopy, Electron, Transmission / Q000379:methods; D008869:Microtubule-Associated Proteins / Q000235:genetics / Q000378:metabolism*; D028361:Mitochondrial Diseases / Q000150:complications / Q000201:enzymology*; D024101:Mitochondrial Proteins / Q000187:drug effects / Q000235:genetics / Q000378:metabolism*; D011180:Postmortem Changes; D055503:Protein Multimerization / Q000187:drug effects / Q000502:physiology
D008869:Microtubule-Associated Proteins; D024101:Mitochondrial Proteins; D001710:Biotin; D020558:GTP Phosphohydrolases; C110768:DNM1L protein, human; D034281:Dynamins; D003545:Cysteine; D004229:Dithiothreitol
J Neurosci. 2004 Oct 20;24(42):9232-9
15496657
Am J Pathol. 2008 Aug;173(2):470-82
18599615
J Biol Chem. 2006 Jan 27;281(4):2177-83
16272155
J Biol Chem. 2004 Aug 20;279(34):35967-74
15208300
Proc Natl Acad Sci U S A. 2004 Nov 9;101(45):15927-32
15509649
J Neurochem. 2005 Feb;92(3):628-36
15659232
Science. 2009 Apr 3;324(5923):102-5
19342591
Proc Natl Acad Sci U S A. 2008 Dec 9;105(49):19318-23
19050078
Apoptosis. 2010 Nov;15(11):1354-63
20177970
J Neurosci. 2001 Jun 15;21(12):4183-7
11404403
FEBS Lett. 2002 Jul 17;523(1-3):171-6
12123827
J Neurochem. 1994 Sep;63(3):910-6
7519665
FEBS Lett. 2002 Aug 14;525(1-3):43-7
12163159
J Biol Chem. 2003 Mar 7;278(10):7743-6
12509422
Neuron. 1992 Jun;8(6):1087-99
1376999
Proc Natl Acad Sci U S A. 2004 Jul 20;101(29):10726-31
15247418
J Cell Biol. 2003 Jan 20;160(2):189-200
12527753
FEBS Lett. 1994 Dec 19;356(2-3):295-8
7805858
Cell. 2006 Oct 20;127(2):383-95
17055438
Neurochem Int. 2009 Feb;54(2):84-8
19114068
Proc Natl Acad Sci U S A. 2008 Oct 14;105(41):15803-8
18838687
Mol Biol Cell. 2001 Aug;12(8):2245-56
11514614
J Neurochem. 2009 May;109 Suppl 1:153-9
19393022
Neurology. 1990 Aug;40(8):1302-3
2166249
Methods Cell Biol. 2008;88:237-56
18617037
J Neurosci. 2005 Jan 19;25(3):672-9
15659604
Nat Rev Neurosci. 2008 Jul;9(7):505-18
18568013
Neurosci Lett. 1995 Jul 7;193(3):201-4
7478183
Neurobiol Dis. 2002 Aug;10(3):258-67
12270688
J Neurosci. 2009 Jul 15;29(28):9090-103
19605646
Hum Mol Genet. 2006 May 1;15(9):1437-49
16551656
EMBO Rep. 2007 Oct;8(10):939-44
17721437
Antioxid Redox Signal. 2009 Mar;11(3):541-54
18715148
Nat Cell Biol. 2001 Feb;3(2):193-7
11175752
Cell. 2006 Jun 30;125(7):1241-52
16814712
Neurosci Lett. 2004 Feb 26;357(1):63-7
15036614
Cell Death Differ. 2007 Mar;14(3):462-71
17053808
Cell Death Differ. 2009 Jun;16(6):899-909
19300456
Neuron. 2004 Feb 5;41(3):351-65
14766175
Methods Enzymol. 2005;404:611-9
16413304
J Cell Sci. 2004 Dec 15;117(Pt 26):6535-46
15572413
J Neurochem. 2002 Jan;80(1):91-100
11796747
Nature. 2004 Jun 24;429(6994):883-91
15190254
Science. 2004 Apr 16;304(5669):448-52
15087549
Cell Death Differ. 2007 Oct;14(10):1721-32
17571083
Trends Neurosci. 2008 Sep;31(9):454-63
18675468
Mol Biol Cell. 2001 Sep;12(9):2894-905
11553726
J Biol Chem. 2007 Apr 13;282(15):11521-9
17301055
J Cell Biol. 2008 Aug 11;182(3):573-85
18695047
Sci STKE. 2001 Jun 12;2001(86):pl1
11752655
Curr Opin Cell Biol. 2003 Dec;15(6):706-16
14644195
EMBO J. 2006 Aug 23;25(16):3900-11
16874299
Mol Cell. 1999 Nov;4(5):815-26
10619028
J Biol Chem. 2007 Dec 21;282(51):37006-15
17965419
EMBO Rep. 2007 Dec;8(12):1088-9; author reply 1089-90
18059302
J Biol Chem. 1999 Jan 29;274(5):2780-5
9915810
J Neuropathol Exp Neurol. 2001 Aug;60(8):759-67
11487050
J Neurosci. 2006 Oct 11;26(41):10480-7
17035532
Arch Neurol. 1988 Aug;45(8):836-40
3395256