PMID- 22125063
OWN - NLM
STAT- MEDLINE
DCOM- 20120316
LR  - 20211203
IS  - 1940-6029 (Electronic)
IS  - 1064-3745 (Linking)
VI  - 821
DP  - 2012
TI  - Detection of cytoplasmic and nuclear functions of mTOR by fractionation.
PG  - 105-24
LID - 10.1007/978-1-61779-430-8_8 [doi]
AB  - Subcellular localization constitutes the environment in which proteins act. It 
      tightly controls access to and availability of different types of molecular 
      interacting partners and is therefore a major determinant of protein function and 
      regulation. Originally thought to be a mere cytoplasmic kinase the mammalian 
      target of rapamycin (mTOR) has recently been localized to various intracellular 
      compartments including the nucleus and specific components of the endomembrane 
      system such as lysosomes. The identification of essential binding partners and 
      the structural and functional partitioning of mTOR into two distinct multiprotein 
      complexes warrant the detailed investigation of the subcellular localization of 
      mTOR as part of mTORC1 and mTORC2. Upon establishment of experimental conditions 
      allowing cytoplasmic/nuclear fractionation at high purity and maximum mTOR 
      complex recovery we have previously shown that the mTOR/raptor complex (mTORC1) 
      is predominantly cytoplasmic whereas the mTOR/rictor complex (mTORC2) is abundant 
      in both compartments. Moreover, the mTORC2 complex components rictor and sin1 are 
      dephosphorylated and dynamically distributed between the cytoplasm and the 
      nucleus upon long-term treatment with the mTOR-inhibitor rapamycin. These 
      findings further demonstrate that the here presented and detailly described 
      fractionation procedure is a valuable tool to study protein localization and 
      cytoplasmic/nuclear protein shuttling in the context of expanding mTOR 
      signalling.
FAU - Rosner, Margit
AU  - Rosner M
AD  - Medical Genetics, Medical University of Vienna, Vienna, Austria.
FAU - Hengstschlager, Markus
AU  - Hengstschlager M
LA  - eng
PT  - Journal Article
PT  - Research Support, Non-U.S. Gov't
PL  - United States
TA  - Methods Mol Biol
JT  - Methods in molecular biology (Clifton, N.J.)
JID - 9214969
RN  - 0 (CRTC2 protein, human)
RN  - 0 (Multiprotein Complexes)
RN  - 0 (Proteins)
RN  - 0 (Transcription Factors)
RN  - EC 2.7.11.1 (Mechanistic Target of Rapamycin Complex 1)
RN  - EC 2.7.11.1 (TOR Serine-Threonine Kinases)
SB  - IM
MH  - Animals
MH  - Cell Fractionation/*methods
MH  - Cell Nucleus/*metabolism
MH  - Cytoplasm/*metabolism
MH  - Cytosol
MH  - Fibroblasts/metabolism
MH  - Genetic Vectors
MH  - Humans
MH  - Mechanistic Target of Rapamycin Complex 1
MH  - Multiprotein Complexes/*metabolism
MH  - Phosphorylation
MH  - Proteins/metabolism
MH  - Signal Transduction
MH  - TOR Serine-Threonine Kinases
MH  - Transcription Factors/metabolism
EDAT- 2011/11/30 06:00
MHDA- 2012/03/17 06:00
CRDT- 2011/11/30 06:00
PHST- 2011/11/30 06:00 [entrez]
PHST- 2011/11/30 06:00 [pubmed]
PHST- 2012/03/17 06:00 [medline]
AID - 10.1007/978-1-61779-430-8_8 [doi]
PST - ppublish
SO  - Methods Mol Biol. 2012;821:105-24. doi: 10.1007/978-1-61779-430-8_8.