[HTML][HTML] ATP and MO25α regulate the conformational state of the STRADα pseudokinase and activation of the LKB1 tumour suppressor

E Zeqiraj, BM Filippi, S Goldie, I Navratilova… - PLoS …, 2009 - journals.plos.org
E Zeqiraj, BM Filippi, S Goldie, I Navratilova, J Boudeau, M Deak, DR Alessi
PLoS biology, 2009journals.plos.org
Pseudokinases lack essential residues for kinase activity, yet are emerging as important
regulators of signal transduction networks. The pseudokinase STRAD activates the LKB1
tumour suppressor by forming a heterotrimeric complex with LKB1 and the scaffolding
protein MO25. Here, we describe the structure of STRADα in complex with MO25α. The
structure reveals an intricate web of interactions between STRADα and MO25α involving the
αC-helix of STRADα, reminiscent of the mechanism by which CDK2 interacts with cyclin A …
Pseudokinases lack essential residues for kinase activity, yet are emerging as important regulators of signal transduction networks. The pseudokinase STRAD activates the LKB1 tumour suppressor by forming a heterotrimeric complex with LKB1 and the scaffolding protein MO25. Here, we describe the structure of STRADα in complex with MO25α. The structure reveals an intricate web of interactions between STRADα and MO25α involving the αC-helix of STRADα, reminiscent of the mechanism by which CDK2 interacts with cyclin A. Surprisingly, STRADα binds ATP and displays a closed conformation and an ordered activation loop, typical of active protein kinases. Inactivity is accounted for by nonconservative substitution of almost all essential catalytic residues. We demonstrate that binding of ATP enhances the affinity of STRADα for MO25α, and conversely, binding of MO25α promotes interaction of STRADα with ATP. Mutagenesis studies reveal that association of STRADα with either ATP or MO25α is essential for LKB1 activation. We conclude that ATP and MO25α cooperate to maintain STRADα in an “active” closed conformation required for LKB1 activation. It has recently been demonstrated that a mutation in human STRADα that truncates a C-terminal region of the pseudokinase domain leads to the polyhydramnios, megalencephaly, symptomatic epilepsy (PMSE) syndrome. We demonstrate this mutation destabilizes STRADα and prevents association with LKB1. In summary, our findings describe one of the first structures of a genuinely inactive pseudokinase. The ability of STRADα to activate LKB1 is dependent on a closed “active” conformation, aided by ATP and MO25α binding. Thus, the function of STRADα is mediated through an active kinase conformation rather than kinase activity. It is possible that other pseudokinases exert their function through nucleotide binding and active conformations.
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