PMID- 22761693
OWN - NLM
STAT- MEDLINE
DCOM- 20130326
LR  - 20211021
IS  - 1932-6203 (Electronic)
IS  - 1932-6203 (Linking)
VI  - 7
IP  - 6
DP  - 2012
TI  - Coaggregation of RNA-binding proteins in a model of TDP-43 proteinopathy with 
      selective RGG motif methylation and a role for RRM1 ubiquitination.
PG  - e38658
LID - 10.1371/journal.pone.0038658 [doi]
LID - e38658
AB  - TAR DNA-binding protein 43 (TDP-43) is a major component within 
      ubiquitin-positive inclusions of a number of neurodegenerative diseases that 
      increasingly are considered as TDP-43 proteinopathies. Identities of other 
      inclusion proteins associated with TDP-43 aggregation remain poorly defined. In 
      this study, we identify and quantitate 35 co-aggregating proteins in the 
      detergent-resistant fraction of HEK-293 cells in which TDP-43 or a particularly 
      aggregate prone variant, TDP-S6, were enriched following overexpression, using 
      stable isotope-labeled (SILAC) internal standards and liquid chromatography 
      coupled to tandem mass spectrometry (LC-MS/MS). We also searched for differential 
      post-translational modification (PTM) sites of ubiquitination. Four sites of 
      ubiquitin conjugation to TDP-43 or TDP-S6 were confirmed by dialkylated 
      GST-TDP-43 external reference peptides, occurring on or near RNA binding motif 
      (RRM) 1. RRM-containing proteins co-enriched in cytoplasmic granular structures 
      in HEK-293 cells and primary motor neurons with insoluble TDP-S6, including 
      cytoplasmic stress granule associated proteins G3BP, PABPC1, and eIF4A1. 
      Proteomic evidence for TDP-43 co-aggregation with paraspeckle markers RBM14, PSF 
      and NonO was also validated by western blot and by immunocytochemistry in HEK-293 
      cells. An increase in peptides from methylated arginine-glycine-glycine (RGG) 
      RNA-binding motifs of FUS/TLS and hnRNPs was found in the detergent-insoluble 
      fraction of TDP-overexpressing cells. Finally, TDP-43 and TDP-S6 
      detergent-insoluble species were reduced by mutagenesis of the identified 
      ubiquitination sites, even following oxidative or proteolytic stress. Together, 
      these findings define some of the aggregation partners of TDP-43, and suggest 
      that TDP-43 ubiquitination influences TDP-43 oligomerization.
FAU - Dammer, Eric B
AU  - Dammer EB
AD  - Department of Human Genetics, Emory University School of Medicine, Atlanta, 
      Georgia, United States of America.
FAU - Fallini, Claudia
AU  - Fallini C
FAU - Gozal, Yair M
AU  - Gozal YM
FAU - Duong, Duc M
AU  - Duong DM
FAU - Rossoll, Wilfried
AU  - Rossoll W
FAU - Xu, Ping
AU  - Xu P
FAU - Lah, James J
AU  - Lah JJ
FAU - Levey, Allan I
AU  - Levey AI
FAU - Peng, Junmin
AU  - Peng J
FAU - Bassell, Gary J
AU  - Bassell GJ
FAU - Seyfried, Nicholas T
AU  - Seyfried NT
LA  - eng
GR  - T32 GM008169/GM/NIGMS NIH HHS/United States
GR  - F32AG038259/AG/NIA NIH HHS/United States
GR  - F32 AG038259/AG/NIA NIH HHS/United States
GR  - F32 AG032848/AG/NIA NIH HHS/United States
GR  - T32 NS007480/NS/NINDS NIH HHS/United States
GR  - NS-007480/NS/NINDS NIH HHS/United States
GR  - P30 NS055077/NS/NINDS NIH HHS/United States
GR  - F32AG032848-02/AG/NIA NIH HHS/United States
GR  - P30NS055077/NS/NINDS NIH HHS/United States
PT  - Journal Article
PT  - Research Support, N.I.H., Extramural
PT  - Research Support, Non-U.S. Gov't
DEP - 20120621
PL  - United States
TA  - PLoS One
JT  - PloS one
JID - 101285081
RN  - 0 (DNA-Binding Proteins)
RN  - 0 (Peptide Fragments)
RN  - 0 (RNA, Messenger)
RN  - 0 (RNA-Binding Proteins)
RN  - 0 (Tumor Suppressor Proteins)
RN  - 94ZLA3W45F (Arginine)
RN  - EC 1.17.4.1 (RRM1 protein, human)
RN  - EC 1.17.4.1 (Ribonucleoside Diphosphate Reductase)
RN  - TE7660XO1C (Glycine)
SB  - IM
MH  - Amino Acid Motifs/*physiology
MH  - Animals
MH  - Arginine/chemistry
MH  - Blotting, Western
MH  - Cell Nucleus/metabolism
MH  - Cytoplasm/metabolism
MH  - DNA-Binding Proteins/genetics/*metabolism
MH  - Glycine/chemistry
MH  - HEK293 Cells
MH  - Humans
MH  - Methylation
MH  - Mice
MH  - Mice, Inbred C57BL
MH  - Motor Neurons/cytology/metabolism
MH  - Peptide Fragments/metabolism
MH  - Protein Multimerization
MH  - Protein Processing, Post-Translational
MH  - Proteomics
MH  - RNA, Messenger/genetics
MH  - RNA-Binding Proteins/genetics/*metabolism
MH  - Real-Time Polymerase Chain Reaction
MH  - Reverse Transcriptase Polymerase Chain Reaction
MH  - Ribonucleoside Diphosphate Reductase
MH  - TDP-43 Proteinopathies/genetics/*metabolism/pathology
MH  - Tandem Mass Spectrometry
MH  - Tumor Suppressor Proteins/genetics/*metabolism
MH  - *Ubiquitination
PMC - PMC3380899
COIS- Competing Interests: The authors have declared that no competing interests exist.
EDAT- 2012/07/05 06:00
MHDA- 2013/03/27 06:00
PMCR- 2012/06/21
CRDT- 2012/07/05 06:00
PHST- 2011/10/17 00:00 [received]
PHST- 2012/05/08 00:00 [accepted]
PHST- 2012/07/05 06:00 [entrez]
PHST- 2012/07/05 06:00 [pubmed]
PHST- 2013/03/27 06:00 [medline]
PHST- 2012/06/21 00:00 [pmc-release]
AID - PONE-D-11-20379 [pii]
AID - 10.1371/journal.pone.0038658 [doi]
PST - ppublish
SO  - PLoS One. 2012;7(6):e38658. doi: 10.1371/journal.pone.0038658. Epub 2012 Jun 21.