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14-3-3 cooperates with LKB1 to regulate the activity and localization of QSK and SIK

Al-Hakim, Abdallah K; Göransson, Olga LU ; Deak, Maria; Toth, Rachel; Campbell, David G; Morrice, Nick A; Prescott, Alan R and Alessi, Dario R (2005) In Journal of Cell Science 118(Pt 23). p.5661-5673
Abstract
The LKB1 tumour suppressor kinase phosphorylates and activates a number of protein kinases belonging to the AMP-activated protein kinase (AMPK) subfamily. We have used a modified tandem affinity purification strategy to identify proteins that interact with AMPKalpha, as well as the twelve AMPK-related kinases that are activated by LKB1. The AMPKbeta and AMPKgamma regulatory subunits were associated with AMPKalpha, but not with any of the AMPK-related kinases, explaining why AMP does not influence the activity of these enzymes. In addition, we identified novel binding partners that interacted with one or more of the AMPK subfamily enzymes, including fat facets/ubiquitin specific protease-9 (USP9), AAA-ATPase-p97, adenine nucleotide... (More)
The LKB1 tumour suppressor kinase phosphorylates and activates a number of protein kinases belonging to the AMP-activated protein kinase (AMPK) subfamily. We have used a modified tandem affinity purification strategy to identify proteins that interact with AMPKalpha, as well as the twelve AMPK-related kinases that are activated by LKB1. The AMPKbeta and AMPKgamma regulatory subunits were associated with AMPKalpha, but not with any of the AMPK-related kinases, explaining why AMP does not influence the activity of these enzymes. In addition, we identified novel binding partners that interacted with one or more of the AMPK subfamily enzymes, including fat facets/ubiquitin specific protease-9 (USP9), AAA-ATPase-p97, adenine nucleotide translocase, protein phosphatase 2A holoenzyme and isoforms of the phospho-protein binding adaptor 14-3-3. Interestingly, the 14-3-3 isoforms bound directly to the T-loop Thr residue of QSK and SIK, after these were phosphorylated by LKB1. Consistent with this, the 14-3-3 isoforms failed to interact with non-phosphorylated QSK and SIK, in LKB1 knockout muscle or in HeLa cells in which LKB1 is not expressed. Moreover, mutation of the T-loop Thr phosphorylated by LKB1, prevented QSK and SIK from interacting with 14-3-3 in vitro. Binding of 14-3-3 to QSK and SIK, enhanced catalytic activity towards the TORC2 protein and the AMARA peptide, and was required for the cytoplasmic localization of SIK and for localization of QSK to punctate structures within the cytoplasm. To our knowledge, this study provides the first example of 14-3-3 binding directly to the T-loop of a protein kinase and influencing its catalytic activity and cellular localization. (Less)
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author
publishing date
type
Contribution to journal
publication status
published
subject
keywords
Mass spectrometry, Cell polarity, AMPK, MARK/Par1, Tandem affinity purification
in
Journal of Cell Science
volume
118
issue
Pt 23
pages
5661 - 5673
publisher
The Company of Biologists Ltd
external identifiers
  • pmid:16306228
  • scopus:30544437191
ISSN
0021-9533
DOI
10.1242/jcs.02670
language
English
LU publication?
no
id
8a8bf920-8cdc-421c-8aee-1facacdd6540 (old id 1132611)
date added to LUP
2008-06-23 15:02:30
date last changed
2017-08-27 04:16:07
@article{8a8bf920-8cdc-421c-8aee-1facacdd6540,
  abstract     = {The LKB1 tumour suppressor kinase phosphorylates and activates a number of protein kinases belonging to the AMP-activated protein kinase (AMPK) subfamily. We have used a modified tandem affinity purification strategy to identify proteins that interact with AMPKalpha, as well as the twelve AMPK-related kinases that are activated by LKB1. The AMPKbeta and AMPKgamma regulatory subunits were associated with AMPKalpha, but not with any of the AMPK-related kinases, explaining why AMP does not influence the activity of these enzymes. In addition, we identified novel binding partners that interacted with one or more of the AMPK subfamily enzymes, including fat facets/ubiquitin specific protease-9 (USP9), AAA-ATPase-p97, adenine nucleotide translocase, protein phosphatase 2A holoenzyme and isoforms of the phospho-protein binding adaptor 14-3-3. Interestingly, the 14-3-3 isoforms bound directly to the T-loop Thr residue of QSK and SIK, after these were phosphorylated by LKB1. Consistent with this, the 14-3-3 isoforms failed to interact with non-phosphorylated QSK and SIK, in LKB1 knockout muscle or in HeLa cells in which LKB1 is not expressed. Moreover, mutation of the T-loop Thr phosphorylated by LKB1, prevented QSK and SIK from interacting with 14-3-3 in vitro. Binding of 14-3-3 to QSK and SIK, enhanced catalytic activity towards the TORC2 protein and the AMARA peptide, and was required for the cytoplasmic localization of SIK and for localization of QSK to punctate structures within the cytoplasm. To our knowledge, this study provides the first example of 14-3-3 binding directly to the T-loop of a protein kinase and influencing its catalytic activity and cellular localization.},
  author       = {Al-Hakim, Abdallah K and Göransson, Olga and Deak, Maria and Toth, Rachel and Campbell, David G and Morrice, Nick A and Prescott, Alan R and Alessi, Dario R},
  issn         = {0021-9533},
  keyword      = {Mass spectrometry,Cell polarity,AMPK,MARK/Par1,Tandem affinity purification},
  language     = {eng},
  number       = {Pt 23},
  pages        = {5661--5673},
  publisher    = {The Company of Biologists Ltd},
  series       = {Journal of Cell Science},
  title        = {14-3-3 cooperates with LKB1 to regulate the activity and localization of QSK and SIK},
  url          = {http://dx.doi.org/10.1242/jcs.02670},
  volume       = {118},
  year         = {2005},
}