Home LiteratureArticle Details
PMID: 21209826 Published · epublish English Journal Article Research Support, N.I.H., Extramural Research Support, Non-U.S. Gov't

The C-terminal TDP-43 fragments have a high aggregation propensity and harm neurons by a dominant-negative mechanism.

PloS one ·Vol. 5 ·No. 12 ·2010-12-31 ·Pages e15878

Yang C, Tan W, Whittle C, Qiu L, Cao L, Akbarian S, Xu Z

Abstract

TAR DNA binding protein 43 KD (TDP-43) is an essential gene that regulates gene transcription, mRNA splicing and stability. In amyotrophic lateral sclerosis (ALS) and frontotemporal dementia (FTD), two fatal neurodegenerative diseases, TDP-43 is fragmented, generating multiple fragments that include the C-terminal fragment of ∼25 KD. The role of these fragments in the pathogenesis of ALS and FTD is not clear. Here we investigated the aggregation propensity in various polypeptide regions of TDP-43 in mammalian cells and the effect of these fragments on cultured neurons. By expressing the full length and various TDP-43 fragments in motor neuron-derived NSC-34 cells and primary neurons, we found that both N- and C-terminal fragments of TDP-43 are prone to aggregate and the C-terminal end of RRM2 region is required, though not sufficient, for aggregation. The aggregation of the TDP-43 fragments can drive co-aggregation with the full-length TDP-43, consequently reducing the nuclear TDP-43. In addition, the TDP-43 fragments can impair neurite growth during neuronal differentiation. Importantly, overexpression of the full-length TDP-43 rescues the neurite growth phenotype whereas knockdown of the endogenous TDP-43 reproduces this phenotype. These results suggest that TDP-43 fragments, particularly the pathologically relevant C-terminal fragments, can impair neuronal differentiation by dominant-negatively interfering with the function of the full length TDP-43, thus playing a role in pathogenesis in ALS and FTD.

MeSH Terms
Animals Cell Differentiation Cell Line, Tumor Cloning, Molecular DNA-Binding Proteins/chemistry Exons Genes, Dominant Genetic Vectors Green Fluorescent Proteins/chemistry Humans Neurons/metabolism Protein Structure, Tertiary RNA Splicing RNA, Messenger/metabolism Transfection
Chemicals
DNA-Binding Proteins RNA, Messenger Green Fluorescent Proteins
Authors & Affiliations
7 authors, click to expand affiliations / ORCID
Yang Chunxing
Department of Biochemistry and Molecular Pharmacology, University of Massachusetts Medical School, Worcester, Massachusetts, United States of America.
Tan Weijia
Whittle Catheryne
Qiu Linghua
Cao Lucheng
Akbarian Schahram
Xu Zuoshang
References (51)
51 references, click to expand
  1. TDP43 depletion rescues aberrant CFTR exon 9 skipping.
    FEBS Lett. 2006 Feb 20;580(5):1339-44 PMID: 16458894
  2. Global analysis of TDP-43 interacting proteins reveals strong association with RNA splicing and translation machinery.
    J Proteome Res. 2010 Feb 5;9(2):1104-20 PMID: 20020773
  3. TDP-43 depletion induces neuronal cell damage through dysregulation of Rho family GTPases.
    J Biol Chem. 2009 Aug 14;284(33):22059-22066 PMID: 19535326
  4. TDP-43 and FUS/TLS: emerging roles in RNA processing and neurodegeneration.
    Hum Mol Genet. 2010 Apr 15;19(R1):R46-64 PMID: 20400460
  5. An RNA polymerase II construct synthesizes short-hairpin RNA with a quantitative indicator and mediates highly efficient RNAi.
    Nucleic Acids Res. 2005 Apr 01;33(6):e62 PMID: 15805121
  6. Characterization of alternative isoforms and inclusion body of the TAR DNA-binding protein-43.
    J Biol Chem. 2010 Jan 1;285(1):608-19 PMID: 19887443
  7. ALS-associated mutations in TDP-43 increase its stability and promote TDP-43 complexes with FUS/TLS.
    Proc Natl Acad Sci U S A. 2010 Jul 27;107(30):13318-23 PMID: 20624952
  8. Transgenic rat model of neurodegeneration caused by mutation in the TDP gene.
    PLoS Genet. 2010 Mar 26;6(3):e1000887 PMID: 20361056
  9. TDP-43 is a developmentally regulated protein essential for early embryonic development.
    J Biol Chem. 2010 Feb 26;285(9):6826-34 PMID: 20040602
  10. Gain and loss of function of ALS-related mutations of TARDBP (TDP-43) cause motor deficits in vivo.
    Hum Mol Genet. 2010 Feb 15;19(4):671-83 PMID: 19959528
  11. Phosphorylated TDP-43 in frontotemporal lobar degeneration and amyotrophic lateral sclerosis.
    Ann Neurol. 2008 Jul;64(1):60-70 PMID: 18546284
  12. A yeast TDP-43 proteinopathy model: Exploring the molecular determinants of TDP-43 aggregation and cellular toxicity.
    Proc Natl Acad Sci U S A. 2008 Apr 29;105(17):6439-44 PMID: 18434538
  13. Cytoplasmic mislocalization of TDP-43 is toxic to neurons and enhanced by a mutation associated with familial amyotrophic lateral sclerosis.
    J Neurosci. 2010 Jan 13;30(2):639-49 PMID: 20071528
  14. TDP-43 mutant transgenic mice develop features of ALS and frontotemporal lobar degeneration.
    Proc Natl Acad Sci U S A. 2009 Nov 3;106(44):18809-14 PMID: 19833869
  15. Usefulness of double gene construct for rapid identification of transgenic mice exhibiting tissue-specific gene expression.
    Mol Reprod Dev. 2001 Dec;60(4):446-56 PMID: 11746955
  16. Deletion of TDP-43 down-regulates Tbc1d1, a gene linked to obesity, and alters body fat metabolism.
    Proc Natl Acad Sci U S A. 2010 Sep 14;107(37):16320-4 PMID: 20660762
  17. Enrichment of C-terminal fragments in TAR DNA-binding protein-43 cytoplasmic inclusions in brain but not in spinal cord of frontotemporal lobar degeneration and amyotrophic lateral sclerosis.
    Am J Pathol. 2008 Jul;173(1):182-94 PMID: 18535185
  18. Progressive motor weakness in transgenic mice expressing human TDP-43.
    Neurobiol Dis. 2010 Nov;40(2):404-14 PMID: 20621187
  19. Pathogenic superoxide dismutase structure, folding, aggregation and turnover.
    Curr Opin Chem Biol. 2006 Apr;10(2):131-8 PMID: 16516535
  20. Expression of TDP-43 C-terminal Fragments in Vitro Recapitulates Pathological Features of TDP-43 Proteinopathies.
    J Biol Chem. 2009 Mar 27;284(13):8516-24 PMID: 19164285
  21. Knockdown of transactive response DNA-binding protein (TDP-43) downregulates histone deacetylase 6.
    EMBO J. 2010 Jan 6;29(1):209-21 PMID: 19910924
  22. TDP43 is a human low molecular weight neurofilament (hNFL) mRNA-binding protein.
    Mol Cell Neurosci. 2007 Jun;35(2):320-7 PMID: 17481916
  23. TDP-43, the signature protein of FTLD-U, is a neuronal activity-responsive factor.
    J Neurochem. 2008 May;105(3):797-806 PMID: 18088371
  24. A guide to choosing fluorescent proteins.
    Nat Methods. 2005 Dec;2(12):905-9 PMID: 16299475
  25. Depletion of TDP 43 overrides the need for exonic and intronic splicing enhancers in the human apoA-II gene.
    Nucleic Acids Res. 2005 Oct 27;33(18):6000-10 PMID: 16254078
  26. Higher order arrangement of the eukaryotic nuclear bodies.
    Proc Natl Acad Sci U S A. 2002 Oct 15;99(21):13583-8 PMID: 12361981
  27. Frontotemporal dementia and amyotrophic lateral sclerosis-associated disease protein TDP-43 promotes dendritic branching.
    Mol Brain. 2009 Sep 25;2:30 PMID: 19781077
  28. Cloning and characterization of a novel cellular protein, TDP-43, that binds to human immunodeficiency virus type 1 TAR DNA sequence motifs.
    J Virol. 1995 Jun;69(6):3584-96 PMID: 7745706
  29. Amyotrophic lateral sclerosis-associated proteins TDP-43 and FUS/TLS function in a common biochemical complex to co-regulate HDAC6 mRNA.
    J Biol Chem. 2010 Oct 29;285(44):34097-105 PMID: 20720006
  30. A construct with fluorescent indicators for conditional expression of miRNA.
    BMC Biotechnol. 2008 Oct 07;8:77 PMID: 18840295
  31. cis-requirement for the maintenance of round spermatid-specific transcription.
    Dev Biol. 2006 Jul 15;295(2):781-90 PMID: 16730344
  32. TARDBP mutations in individuals with sporadic and familial amyotrophic lateral sclerosis.
    Nat Genet. 2008 May;40(5):572-4 PMID: 18372902
  33. TDP-43, a neuro-pathosignature factor, is essential for early mouse embryogenesis.
    Genesis. 2010 Jan;48(1):56-62 PMID: 20014337
  34. Characterization and functional implications of the RNA binding properties of nuclear factor TDP-43, a novel splicing regulator of CFTR exon 9.
    J Biol Chem. 2001 Sep 28;276(39):36337-43 PMID: 11470789
  35. Progranulin mediates caspase-dependent cleavage of TAR DNA binding protein-43.
    J Neurosci. 2007 Sep 26;27(39):10530-4 PMID: 17898224
  36. TDP-43 transgenic mice develop spastic paralysis and neuronal inclusions characteristic of ALS and frontotemporal lobar degeneration.
    Proc Natl Acad Sci U S A. 2010 Feb 23;107(8):3858-63 PMID: 20133711
  37. Multiple roles of TDP-43 in gene expression, splicing regulation, and human disease.
    Front Biosci. 2008 Jan 01;13:867-78 PMID: 17981595
  38. Proteolytic processing of TAR DNA binding protein-43 by caspases produces C-terminal fragments with disease defining properties independent of progranulin.
    J Neurochem. 2009 Aug;110(3):1082-94 PMID: 19522733
  39. A Drosophila model for TDP-43 proteinopathy.
    Proc Natl Acad Sci U S A. 2010 Feb 16;107(7):3169-74 PMID: 20133767
  40. Phosphorylated and ubiquitinated TDP-43 pathological inclusions in ALS and FTLD-U are recapitulated in SH-SY5Y cells.
    FEBS Lett. 2009 Jan 22;583(2):394-400 PMID: 19111550
  41. Structural diversity and functional implications of the eukaryotic TDP gene family.
    Genomics. 2004 Jan;83(1):130-9 PMID: 14667816
  42. Nuclear factor TDP-43 and SR proteins promote in vitro and in vivo CFTR exon 9 skipping.
    EMBO J. 2001 Apr 2;20(7):1774-84 PMID: 11285240
  43. TDP-43 is intrinsically aggregation-prone, and amyotrophic lateral sclerosis-linked mutations accelerate aggregation and increase toxicity.
    J Biol Chem. 2009 Jul 24;284(30):20329-39 PMID: 19465477
  44. Truncation and pathogenic mutations facilitate the formation of intracellular aggregates of TDP-43.
    Hum Mol Genet. 2009 Sep 15;18(18):3353-64 PMID: 19515851
  45. Disturbance of nuclear and cytoplasmic TAR DNA-binding protein (TDP-43) induces disease-like redistribution, sequestration, and aggregate formation.
    J Biol Chem. 2008 May 9;283(19):13302-9 PMID: 18305110
  46. Ubiquitinated TDP-43 in frontotemporal lobar degeneration and amyotrophic lateral sclerosis.
    Science. 2006 Oct 6;314(5796):130-3 PMID: 17023659
  47. Structural insights into TDP-43 in nucleic-acid binding and domain interactions.
    Nucleic Acids Res. 2009 Apr;37(6):1799-808 PMID: 19174564
  48. Neuroblastoma x spinal cord (NSC) hybrid cell lines resemble developing motor neurons.
    Dev Dyn. 1992 Jul;194(3):209-21 PMID: 1467557
  49. Aberrant cleavage of TDP-43 enhances aggregation and cellular toxicity.
    Proc Natl Acad Sci U S A. 2009 May 5;106(18):7607-12 PMID: 19383787
  50. Wild-type human TDP-43 expression causes TDP-43 phosphorylation, mitochondrial aggregation, motor deficits, and early mortality in transgenic mice.
    J Neurosci. 2010 Aug 11;30(32):10851-9 PMID: 20702714
  51. Loss of murine TDP-43 disrupts motor function and plays an essential role in embryogenesis.
    Acta Neuropathol. 2010 Apr;119(4):409-19 PMID: 20198480
Article Info
Journal
PloS one
Abbr.
PLoS One
ISSN
1932-6203
Published
2010-12-31
Epub
2010-00-31
Pages
e15878
Language
English
Region
United States
NLM ID
101285081
PMCID
PMC3013128
Subset
IM
Grants
NINDS NIH HHS · R01 NS048145 · United States
NINDS NIH HHS · R21 NS062230 · United States
Analysis Services
Analysis Services

Contact

No. 2 Wenbo Road, Zhangqiu District, Jinan, Shandong

Qilu Normal University · Genelibs Bioinformatics Lab

750 Shunhua Rd, Jinan

2F, Bldg F, University Science Park

Tel: 0531-88819269

WeChat Official Account

Follow our WeChat subscription account for real-time updates and the latest in medical and biological research.


Business Email

E-mail: product@genelibs.com