PMID- 28174230
OWN - NLM
STAT- MEDLINE
DCOM- 20170627
LR  - 20211204
IS  - 1535-9484 (Electronic)
IS  - 1535-9476 (Print)
IS  - 1535-9476 (Linking)
VI  - 16
IP  - 4
DP  - 2017 Apr
TI  - Clustered, Regularly Interspaced Short Palindromic Repeats (CRISPR)/Cas9-coupled 
      Affinity Purification/Mass Spectrometry Analysis Revealed a Novel Role of 
      Neurofibromin in mTOR Signaling.
PG  - 594-607
LID - 10.1074/mcp.M116.064543 [doi]
AB  - Neurofibromin (NF1) is a well known tumor suppressor that is commonly mutated in 
      cancer patients. It physically interacts with RAS and negatively regulates RAS 
      GTPase activity. Despite the importance of NF1 in cancer, a high quality 
      endogenous NF1 interactome has yet to be established. In this study, we combined 
      clustered, regularly interspaced short palindromic repeats (CRISPR)/Cas9-mediated 
      gene knock-out technology with affinity purification using antibodies against 
      endogenous proteins, followed by mass spectrometry analysis, to sensitively and 
      accurately detect NF1 protein-protein interactions in unaltered in vivo settings. 
      Using this system, we analyzed endogenous NF1-associated protein complexes and 
      identified 49 high-confidence candidate interaction proteins, including RAS and 
      other functionally relevant proteins. Through functional validation, we found 
      that NF1 negatively regulates mechanistic target of rapamycin signaling (mTOR) in 
      a LAMTOR1-dependent manner. In addition, the cell growth and survival of 
      NF1-deficient cells have become dependent on hyperactivation of the mTOR pathway, 
      and the tumorigenic properties of these cells have become dependent on LAMTOR1. 
      Taken together, our findings may provide novel insights into therapeutic 
      approaches targeting NF1-deficient tumors.
CI  - (c) 2017 by The American Society for Biochemistry and Molecular Biology, Inc.
FAU - Li, Xu
AU  - Li X
AD  - From the double daggerDepartment of Experimental Radiation Oncology, University of Texas M.D. 
      Anderson Cancer Center, Houston, Texas 77030.
FAU - Gao, Min
AU  - Gao M
AD  - From the double daggerDepartment of Experimental Radiation Oncology, University of Texas M.D. 
      Anderson Cancer Center, Houston, Texas 77030.
FAU - Choi, Jong Min
AU  - Choi JM
AD  -  ||Department of Molecular and Cellular Biology, Baylor College of Medicine, 
      Houston, Texas 77030.
FAU - Kim, Beom-Jun
AU  - Kim BJ
AD  -  ||Department of Molecular and Cellular Biology, Baylor College of Medicine, 
      Houston, Texas 77030.
FAU - Zhou, Mao-Tian
AU  - Zhou MT
AD  - From the double daggerDepartment of Experimental Radiation Oncology, University of Texas M.D. 
      Anderson Cancer Center, Houston, Texas 77030.
FAU - Chen, Zhen
AU  - Chen Z
AD  - From the double daggerDepartment of Experimental Radiation Oncology, University of Texas M.D. 
      Anderson Cancer Center, Houston, Texas 77030.
FAU - Jain, Antrix N
AU  - Jain AN
AD  -  ||Department of Molecular and Cellular Biology, Baylor College of Medicine, 
      Houston, Texas 77030.
FAU - Jung, Sung Yun
AU  - Jung SY
AD  -  ||Department of Molecular and Cellular Biology, Baylor College of Medicine, 
      Houston, Texas 77030.
FAU - Yuan, Jingsong
AU  - Yuan J
AD  - **Department of Radiation Oncology, Center for Radiological Research, Columbia 
      University, New York, New York 10032.
FAU - Wang, Wenqi
AU  - Wang W
AD  - double daggerdouble daggerDepartment of Developmental and Cell Biology, University of California at 
      Irvine, Irvine, California 92697 jchen8@mdanderson.org yiw@bcm.edu 
      wenqiw6@uci.edu.
FAU - Wang, Yi
AU  - Wang Y
AD  -  ||Department of Molecular and Cellular Biology, Baylor College of Medicine, 
      Houston, Texas 77030; jchen8@mdanderson.org yiw@bcm.edu wenqiw6@uci.edu.
FAU - Chen, Junjie
AU  - Chen J
AD  - From the double daggerDepartment of Experimental Radiation Oncology, University of Texas M.D. 
      Anderson Cancer Center, Houston, Texas 77030; jchen8@mdanderson.org yiw@bcm.edu 
      wenqiw6@uci.edu.
LA  - eng
GR  - P30 CA016672/CA/NCI NIH HHS/United States
PT  - Journal Article
DEP - 20170207
PL  - United States
TA  - Mol Cell Proteomics
JT  - Molecular & cellular proteomics : MCP
JID - 101125647
RN  - 0 (Carrier Proteins)
RN  - 0 (Intracellular Signaling Peptides and Proteins)
RN  - 0 (LAMTOR1 protein, human)
RN  - 0 (Neurofibromin 1)
RN  - EC 2.7.1.1 (MTOR protein, human)
RN  - EC 2.7.11.1 (TOR Serine-Threonine Kinases)
SB  - IM
MH  - CRISPR-Cas Systems
MH  - Carrier Proteins/genetics/*metabolism
MH  - Cell Proliferation
MH  - Cell Survival
MH  - Gene Editing/*methods
MH  - HEK293 Cells
MH  - HeLa Cells
MH  - Humans
MH  - Intracellular Signaling Peptides and Proteins
MH  - Mass Spectrometry
MH  - Neoplasms/genetics/*metabolism
MH  - Neurofibromin 1/genetics/*metabolism
MH  - Protein Interaction Maps
MH  - Proteomics/*methods
MH  - Signal Transduction
MH  - TOR Serine-Threonine Kinases/genetics/*metabolism
PMC - PMC5383781
COIS- The authors declare that they have no conflicts of interest with the contents of 
      this article
EDAT- 2017/02/09 06:00
MHDA- 2017/06/28 06:00
PMCR- 2018/04/01
CRDT- 2017/02/09 06:00
PHST- 2016/10/06 00:00 [received]
PHST- 2017/01/25 00:00 [revised]
PHST- 2017/02/09 06:00 [pubmed]
PHST- 2017/06/28 06:00 [medline]
PHST- 2017/02/09 06:00 [entrez]
PHST- 2018/04/01 00:00 [pmc-release]
AID - S1535-9476(20)32407-5 [pii]
AID - M116.064543 [pii]
AID - 10.1074/mcp.M116.064543 [doi]
PST - ppublish
SO  - Mol Cell Proteomics. 2017 Apr;16(4):594-607. doi: 10.1074/mcp.M116.064543. Epub 
      2017 Feb 7.