title: Aberrant assembly of RNA recognition motif 1 links to pathogenic conversion of TAR DNA-binding protein of 43 kDa (TDP-43).
abstract: Aggregation of TAR DNA-binding protein of 43 kDa (TDP-43) is a pathological signature of amyotrophic lateral sclerosis (ALS). Although accumulating evidence suggests the involvement of RNA recognition motifs (RRMs) in TDP-43 proteinopathy, it remains unclear how native TDP-43 is converted to pathogenic forms. To elucidate the role of homeostasis of RRM1 structure in ALS pathogenesis, conformations of RRM1 under high pressure were monitored by NMR. We first found that RRM1 was prone to aggregation and had three regions showing stable chemical shifts during misfolding. Moreover, mass spectrometric analysis of aggregated RRM1 revealed that one of the regions was located on protease-resistant β-strands containing two cysteines (Cys-173 and Cys-175), indicating that this region served as a core assembly interface in RRM1 aggregation. Although a fraction of RRM1 aggregates comprised disulfide-bonded oligomers, the substitution of cysteine(s) to serine(s) (C/S) resulted in unexpected acceleration of amyloid fibrils of RRM1 and disulfide-independent aggregate formation of full-length TDP-43. Notably, TDP-43 aggregates with RRM1-C/S required the C terminus, and replicated cytopathologies of ALS, including mislocalization, impaired RNA splicing, ubiquitination, phosphorylation, and motor neuron toxicity. Furthermore, RRM1-C/S accentuated inclusions of familial ALS-linked TDP-43 mutants in the C terminus. The relevance of RRM1-C/S-induced TDP-43 aggregates in ALS pathogenesis was verified by immunolabeling of inclusions of ALS patients and cultured cells overexpressing the RRM1-C/S TDP-43 with antibody targeting misfolding-relevant regions. Our results indicate that cysteines in RRM1 crucially govern the conformation of TDP-43, and aberrant self-assembly of RRM1 at amyloidogenic regions contributes to pathogenic conversion of TDP-43 in ALS.
authors.affiliation: Molecular Neuroscience Research Center, Shiga University of Medical Science, Otsu 520-2192, Japan.
authors.forename: Akemi
authors.initials: A
authors.lastname: Shodai
authors.forename: Toshifumi
authors.initials: T
authors.lastname: Morimura
authors.forename: Akemi
authors.initials: A
authors.lastname: Ido
authors.forename: Tsukasa
authors.initials: T
authors.lastname: Uchida
authors.forename: Takashi
authors.initials: T
authors.lastname: Ayaki
authors.forename: Rina
authors.initials: R
authors.lastname: Takahashi
authors.forename: Soichiro
authors.initials: S
authors.lastname: Kitazawa
authors.forename: Sakura
authors.initials: S
authors.lastname: Suzuki
authors.forename: Mikako
authors.initials: M
authors.lastname: Shirouzu
authors.forename: Takanori
authors.initials: T
authors.lastname: Kigawa
authors.forename: Yutaka
authors.initials: Y
authors.lastname: Muto
authors.forename: Shigeyuki
authors.initials: S
authors.lastname: Yokoyama
authors.forename: Ryosuke
authors.initials: R
authors.lastname: Takahashi
authors.forename: Ryo
authors.initials: R
authors.lastname: Kitahara
authors.forename: Hidefumi
authors.initials: H
authors.lastname: Ito
authors.forename: Noriko
authors.initials: N
authors.lastname: Fujiwara
authors.forename: Makoto
authors.initials: M
authors.lastname: Urushitani
mesh_terms: D020816:Amino Acid Motifs; D000682:Amyloid / Q000737:chemistry / Q000378:metabolism; D000690:Amyotrophic Lateral Sclerosis / Q000378:metabolism / Q000473:pathology; D004268:DNA-Binding Proteins / Q000737:chemistry / Q000378:metabolism; D005260:Female; D057809:HEK293 Cells; D006801:Humans; D045586:Intranuclear Inclusion Bodies / Q000378:metabolism / Q000473:pathology; D009682:Magnetic Resonance Spectroscopy; D008297:Male; D009474:Neurons / Q000378:metabolism / Q000473:pathology; D017510:Protein Folding; D020836:Protein Structure, Quaternary; D017434:Protein Structure, Tertiary; D012326:RNA Splicing; D054875:Ubiquitination
chemical_list: D000682:Amyloid; D004268:DNA-Binding Proteins
keywords: Amyotrophic Lateral Sclerosis (Lou Gehrig's Disease); Cell Biology; Protein Chemical Modification; Protein Misfolding; Structural Biology; TAR DNA-binding Protein 43 kDa
references.citation: Biochem J. 1999 Aug 15;342 ( Pt 1):119-23
references.pmid: 10432308
references.citation: J Neurosci Res. 2010 Mar;88(4):784-97
references.pmid: 19798749
references.citation: Nat Genet. 2008 May;40(5):572-4
references.pmid: 18372902
references.citation: Biochem Biophys Res Commun. 2012 Aug 24;425(2):219-24
references.pmid: 22835933
references.citation: EMBO J. 2012 Mar 7;31(5):1241-52
references.pmid: 22193716
references.citation: Int J Cell Biol. 2012;2012:463756
references.pmid: 22956959
references.citation: Am J Hum Genet. 2004 Jun;74(6):1322-5
references.pmid: 15195661
references.citation: Hum Mol Genet. 2010 Aug 15;19(16):3206-18
references.pmid: 20530643
references.citation: Mol Cell Neurosci. 2011 Jul;47(3):167-80
references.pmid: 21421050
references.citation: Hum Mol Genet. 2011 May 15;20(10):1952-65
references.pmid: 21355045
references.citation: J Neurochem. 2013 Jul;126(2):288-300
references.pmid: 23398327
references.citation: J Biol Chem. 2012 Mar 2;287(10):7512-25
references.pmid: 22235134
references.citation: Ann Neurol. 2008 Jul;64(1):60-70
references.pmid: 18546284
references.citation: J Neurosci Res. 2008 Feb 15;86(3):553-65
references.pmid: 17896795
references.citation: J Biol Chem. 2011 May 27;286(21):18664-72
references.pmid: 21454603
references.citation: Am J Pathol. 2008 Jul;173(1):182-94
references.pmid: 18535185
references.citation: FASEB J. 2011 Jul;25(7):2344-53
references.pmid: 21450909
references.citation: Neurobiol Dis. 2012 Mar;45(3):862-70
references.pmid: 22198567
references.citation: PLoS One. 2012;7(12):e52776
references.pmid: 23300771
references.citation: J Biol Chem. 2011 May 27;286(21):18845-55
references.pmid: 21454607
references.citation: Biochem Biophys Res Commun. 2006 Dec 22;351(3):602-11
references.pmid: 17084815
references.citation: Am J Pathol. 2012 Jan;180(1):293-302
references.pmid: 22067910
references.citation: Proc Natl Acad Sci U S A. 2007 Jul 17;104(29):11921-6
references.pmid: 17616582
references.citation: J Biol Chem. 2001 Sep 28;276(39):36337-43
references.pmid: 11470789
references.citation: J Biol Chem. 2012 Jun 29;287(27):23079-94
references.pmid: 22563080
references.citation: Nat Neurosci. 2011 Apr;14(4):452-8
references.pmid: 21358640
references.citation: J Mol Biol. 2006 Oct 20;363(2):395-404
references.pmid: 16979187
references.citation: Nat Neurosci. 2011 Apr;14(4):459-68
references.pmid: 21358643
references.citation: FEBS Lett. 2009 Jan 22;583(2):394-400
references.pmid: 19111550
references.citation: J Cell Sci. 2008 Nov 15;121(Pt 22):3778-85
references.pmid: 18957508
references.citation: Hum Mol Genet. 2009 Oct 15;18(R2):R156-62
references.pmid: 19808791
references.citation: Trends Mol Med. 2011 Nov;17(11):659-67
references.pmid: 21783422
references.citation: Brain. 2011 Sep;134(Pt 9):2610-26
references.pmid: 21752789
references.citation: J Biol Chem. 2007 Dec 7;282(49):35933-44
references.pmid: 17913710
references.citation: J Clin Invest. 2011 Feb;121(2):726-38
references.pmid: 21206091
references.citation: Chem Rev. 2006 May;106(5):1814-35
references.pmid: 16683756
references.citation: J Exp Med. 2011 Nov 21;208(12):2429-47
references.pmid: 22084410
references.citation: Neuropathol Appl Neurobiol. 2010 Apr;36(2):97-112
references.pmid: 20202122
references.citation: Annu Rev Biochem. 2006;75:333-66
references.pmid: 16756495
references.citation: Curr Opin Neurol. 2012 Dec;25(6):701-7
references.pmid: 23041957
references.citation: Biochim Biophys Acta. 2006 Mar;1764(3):546-51
references.pmid: 16298177
references.citation: Biochem Pharmacol. 2006 Feb 28;71(5):551-64
references.pmid: 16337153
references.citation: J Biol Chem. 2008 May 9;283(19):13302-9
references.pmid: 18305110
references.citation: Science. 2006 Oct 6;314(5796):130-3
references.pmid: 17023659
references.citation: Nucleic Acids Res. 2009 Apr;37(6):1799-808
references.pmid: 19174564