title: Analysis of the LKB1-STRAD-MO25 complex.
abstract: Mutations in the LKB1 tumour suppressor threonine kinase cause the inherited Peutz-Jeghers cancer syndrome and are also observed in some sporadic cancers. Recent work indicates that LKB1 exerts effects on metabolism, polarity and proliferation by phosphorylating and activating protein kinases belonging to the AMPK subfamily. In vivo, LKB1 forms a complex with STRAD, an inactive pseudokinase, and MO25, an armadillo repeat scaffolding-like protein. Binding of LKB1 to STRAD-MO25 activates LKB1 and re-localises it from the nucleus to the cytoplasm. To learn more about the inherent properties of the LKB1-STRAD-MO25 complex, we first investigated the activity of 34 point mutants of LKB1 found in human cancers and their ability to interact with STRAD and MO25. Interestingly, 12 of these mutants failed to interact with STRAD-MO25. Performing mutagenesis analysis, we defined two binding sites located on opposite surfaces of MO25alpha, which are required for the assembly of MO25alpha into a complex with STRADalpha and LKB1. In addition, we demonstrate that LKB1 does not require phosphorylation of its own T-loop to be activated by STRADalpha-MO25alpha, and discuss the possibility that this unusual mechanism of regulation arises from LKB1 functioning as an upstream kinase. Finally, we establish that STRADalpha, despite being catalytically inactive, is still capable of binding ATP with high affinity, but that this is not required for activation of LKB1. Taken together, our findings reinforce the functional importance of the binding of LKB1 to STRAD, and provide a greater understanding of the mechanism by which LKB1 is regulated and activated through its interaction with STRAD and MO25.
authors.affiliation: MRC Protein Phosphorylation Unit, MSI/WTB complex, University of Dundee, Dow Street, Dundee, DD1 5EH, Scotland. d.r.alessi@dundee.ac.uk
authors.forename: Jérôme
authors.initials: J
authors.lastname: Boudeau
authors.forename: John W
authors.initials: JW
authors.lastname: Scott
authors.forename: Nicoletta
authors.initials: N
authors.lastname: Resta
authors.forename: Maria
authors.initials: M
authors.lastname: Deak
authors.forename: Agnieszka
authors.initials: A
authors.lastname: Kieloch
authors.forename: David
authors.initials: D
authors.lastname: Komander
authors.forename: D Grahame
authors.initials: DG
authors.lastname: Hardie
authors.forename: Alan R
authors.initials: AR
authors.lastname: Prescott
authors.forename: Daan M F
authors.initials: DM
authors.lastname: van Aalten
authors.forename: Dario R
authors.initials: DR
authors.lastname: Alessi
mesh_terms: D000091162:AMP-Activated Protein Kinase Kinases; D033942:Adaptor Proteins, Vesicular Transport / Q000737:chemistry / Q000378:metabolism*; D000255:Adenosine Triphosphate / Q000378:metabolism; D000595:Amino Acid Sequence; D051186:Armadillo Domain Proteins; D001665:Binding Sites; D020134:Catalytic Domain; D002460:Cell Line; D002467:Cell Nucleus / Q000378:metabolism; D003593:Cytoplasm / Q000378:metabolism; D005786:Gene Expression Regulation; D005982:Glutathione Transferase / Q000378:metabolism; D006367:HeLa Cells; D006801:Humans; D015151:Immunoblotting; D008958:Models, Molecular; D008969:Molecular Sequence Data; D010580:Peutz-Jeghers Syndrome / Q000235:genetics / Q000378:metabolism; D017354:Point Mutation; D017346:Protein Serine-Threonine Kinases / Q000737:chemistry / Q000235:genetics / Q000378:metabolism*; D017434:Protein Structure, Tertiary; D011993:Recombinant Fusion Proteins / Q000378:metabolism; D017386:Sequence Homology, Amino Acid; D015534:Trans-Activators
chemical_list: D033942:Adaptor Proteins, Vesicular Transport; D051186:Armadillo Domain Proteins; D011993:Recombinant Fusion Proteins; C476400:STRADA protein, human; D015534:Trans-Activators; D000255:Adenosine Triphosphate; D005982:Glutathione Transferase; D017346:Protein Serine-Threonine Kinases; C109790:STK11 protein, human; D000091162:AMP-Activated Protein Kinase Kinases