PMID- 23358504
OWN - NLM
STAT- MEDLINE
DCOM- 20131209
LR  - 20211021
IS  - 1535-9484 (Electronic)
IS  - 1535-9476 (Print)
IS  - 1535-9476 (Linking)
VI  - 12
IP  - 5
DP  - 2013 May
TI  - Integrated proteomics identified novel activation of dynein IC2-GR-COX-1 
      signaling in neurofibromatosis type I (NF1) disease model cells.
PG  - 1377-94
LID - 10.1074/mcp.M112.024802 [doi]
AB  - Neurofibromatosis type 1 (NF1) tumor suppressor gene product, neurofibromin, 
      functions in part as a Ras-GAP, and though its loss is implicated in the neuronal 
      abnormality of NF1 patients, its precise cellular function remains unclear. To 
      study the molecular mechanism of NF1 pathogenesis, we prepared NF1 gene knockdown 
      (KD) PC12 cells, as a NF1 disease model, and analyzed their molecular (gene and 
      protein) expression profiles with a unique integrated proteomics approach, 
      comprising iTRAQ, 2D-DIGE, and DNA microarrays, using an integrated protein and 
      gene expression analysis chart (iPEACH). In NF1-KD PC12 cells showing abnormal 
      neuronal differentiation after NGF treatment, of 3198 molecules quantitatively 
      identified and listed in iPEACH, 97 molecules continuously up- or down-regulated 
      over time were extracted. Pathway and network analysis further revealed 
      overrepresentation of calcium signaling and transcriptional regulation by 
      glucocorticoid receptor (GR) in the up-regulated protein set, whereas nerve 
      system development was overrepresented in the down-regulated protein set. The 
      novel up-regulated network we discovered, "dynein IC2-GR-COX-1 signaling," was 
      then examined in NF1-KD cells. Validation studies confirmed that NF1 knockdown 
      induces altered splicing and phosphorylation patterns of dynein IC2 isomers, 
      up-regulation and accumulation of nuclear GR, and increased COX-1 expression in 
      NGF-treated cells. Moreover, the neurite retraction phenotype observed in NF1-KD 
      cells was significantly recovered by knockdown of the dynein IC2-C isoform and 
      COX-1. In addition, dynein IC2 siRNA significantly inhibited nuclear 
      translocation and accumulation of GR and up-regulation of COX-1 expression. These 
      results suggest that dynein IC2 up-regulates GR nuclear translocation and 
      accumulation, and subsequently causes increased COX-1 expression, in this NF1 
      disease model. Our integrated proteomics strategy, which combines multiple 
      approaches, demonstrates that NF1-related neural abnormalities are, in part, 
      caused by up-regulation of dynein IC2-GR-COX-1 signaling, which may be a novel 
      therapeutic target for NF1.
FAU - Hirayama, Mio
AU  - Hirayama M
AD  - Department of Tumor Genetics and Biology, Graduate school of Medical Sciences, 
      Kumamoto University, 1-1-1 Honjo, Chuo-ku, Kumamoto 860-8556, Japan.
FAU - Kobayashi, Daiki
AU  - Kobayashi D
FAU - Mizuguchi, Souhei
AU  - Mizuguchi S
FAU - Morikawa, Takashi
AU  - Morikawa T
FAU - Nagayama, Megumi
AU  - Nagayama M
FAU - Midorikawa, Uichi
AU  - Midorikawa U
FAU - Wilson, Masayo M
AU  - Wilson MM
FAU - Nambu, Akiko N
AU  - Nambu AN
FAU - Yoshizawa, Akiyasu C
AU  - Yoshizawa AC
FAU - Kawano, Shin
AU  - Kawano S
FAU - Araki, Norie
AU  - Araki N
LA  - eng
PT  - Journal Article
PT  - Research Support, Non-U.S. Gov't
DEP - 20130128
PL  - United States
TA  - Mol Cell Proteomics
JT  - Molecular & cellular proteomics : MCP
JID - 101125647
RN  - 0 (Membrane Proteins)
RN  - 0 (Neurofibromin 1)
RN  - 0 (Proteome)
RN  - 0 (Receptors, Glucocorticoid)
RN  - 9061-61-4 (Nerve Growth Factor)
RN  - EC 1.14.99.1 (Cyclooxygenase 1)
RN  - EC 1.14.99.1 (Ptgs1 protein, rat)
RN  - EC 3.6.4.2 (Cytoplasmic Dyneins)
RN  - EC 3.6.4.2 (Dync1i2 protein, rat)
SB  - IM
MH  - Active Transport, Cell Nucleus
MH  - Animals
MH  - Cyclooxygenase 1/genetics/*metabolism
MH  - Cytoplasmic Dyneins/genetics/*metabolism
MH  - Gene Regulatory Networks
MH  - Membrane Proteins/genetics/*metabolism
MH  - Nerve Growth Factor/physiology
MH  - Neurites/metabolism
MH  - Neurofibromatosis 1/metabolism
MH  - Neurofibromin 1/genetics/metabolism
MH  - Oligonucleotide Array Sequence Analysis
MH  - PC12 Cells
MH  - Phosphorylation
MH  - Protein Processing, Post-Translational
MH  - Proteome/genetics/metabolism
MH  - Proteomics
MH  - RNA Splicing
MH  - Rats
MH  - Receptors, Glucocorticoid/genetics/*metabolism
MH  - *Signal Transduction
MH  - Transcriptome
MH  - Up-Regulation
PMC - PMC3650346
EDAT- 2013/01/30 06:00
MHDA- 2013/12/16 06:00
PMCR- 2014/05/01
CRDT- 2013/01/30 06:00
PHST- 2013/01/30 06:00 [entrez]
PHST- 2013/01/30 06:00 [pubmed]
PHST- 2013/12/16 06:00 [medline]
PHST- 2014/05/01 00:00 [pmc-release]
AID - S1535-9476(20)31118-X [pii]
AID - M112.024802 [pii]
AID - 10.1074/mcp.M112.024802 [doi]
PST - ppublish
SO  - Mol Cell Proteomics. 2013 May;12(5):1377-94. doi: 10.1074/mcp.M112.024802. Epub 
      2013 Jan 28.