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Cited by 129PI3K/AKT/mTOR signaling in cancerProtein Kinase Regulation and GTPase SignalingMetabolism, Diabetes, and Cancer

Regulation of lipid binding underlies the activation mechanism of class IA PI3-kinases

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Hon, W-C, Berndt, A, Williams, R L·2012·Oncogene
DOI 10.1038/onc.2011.532PMC PMC3378484
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Abstract

Somatic missense mutations in PIK3CA, which encodes the p110α catalytic subunit of phosphoinositide 3-kinases, occur frequently in human cancers. Activating mutations spread across multiple domains, some of which are located at inhibitory contact sites formed with the regulatory subunit p85α. PIK3R1, which encodes p85α, also has activating somatic mutations. We find a strong correlation between lipid kinase and lipid-binding activities for both wild-type (WT) and a representative set of oncogenic mutant complexes of p110α/p85α. Lipid binding involves both electrostatic and hydrophobic interactions. Activation caused by a phosphorylated receptor tyrosine kinase (RTK) peptide binding to the p85α N-terminal SH2 domain (nSH2) induces lipid binding. This depends on the polybasic activation loop as well as a conserved hydrophobic motif in the C-terminal region of the kinase domain. The hotspot E545K mutant largely mimics the activated WT p110α. It shows the highest basal activity and lipid binding, and is not significantly activated by an RTK phosphopeptide. Both the hotspot H1047R mutant and rare mutations (C420R, M1043I, H1047L, G1049R and p85α-N564D) also show increased basal kinase activities and lipid binding. However, their activities are further enhanced by an RTK phosphopeptide to levels markedly exceeding that of activated WT p110α. Phosphopeptide binding to p110β/p85α and p110δ/p85α complexes also induces their lipid binding. We present a crystal structure of WT p110α complexed with the p85α inter-SH2 domain and the inhibitor PIK-108. Additional to the ATP-binding pocket, an unexpected, second PIK-108 binding site is observed in the kinase C-lobe. We show a global conformational change in p110α consistent with allosteric regulation of the kinase domain by nSH2. These findings broaden our understanding of the differential biological outputs exhibited by distinct types of mutations regarding growth factor dependence, and suggest a two-tier classification scheme relating p110α and p85α mutations with signalling potential.

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  • PI3K activity in the p110α-p85 heterodimer is inhibited by intra- and inter-subunit contacts that become disrupted following engagement of p85 SH2 domains with phosphorylated tyrosine residues in RTK/adapter proteins, and activation favours interaction with the plasma membrane.

    “In this case, the PI3K activity is inhibited by a combination of intra- and inter-subunit contacts that become disrupted following the engagement of the SH2 domains, present in p85, with the phosphorylated tyrosine residues in RTK/adapter proteins in stimulated cells; the activation also favours the interaction with the plasma membrane [74–76].”

    From Phosphatidylinositol(4,5)bisphosphate: diverse functions at the plasma membrane (2020) · cites this paper as [76]

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Authors
Hon, W.-C., Berndt, A., & Williams, R. L.
Affiliations
MRC Laboratory of Molecular Biology (GB)
Corresponding author
Williams, R L
Venue
Oncogene
Publisher
Springer Science and Business Media LLC
Published
2011-11-28
Volume
31
Issue
32
Pages
3655-3666
ISSN
  • 0950-9232 (ISSN-L)
  • 1476-5594
DOI
10.1038/onc.2011.532
PMC
PMC3378484
OpenAlex
W2102005789
Cited by
129
References
2
Citations by year
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FWCI
3.25
Type
journal article
Language
en
Open access
Licence
Locations
  • Oncogene (publishedVersion)
  • PubMed (publishedVersion)
  • Europe PMC (PubMed Central) (acceptedVersion)
  • PubMed Central (acceptedVersion)
Topics
  • Life Sciences › Biochemistry, Genetics and Molecular Biology › Molecular Biology › PI3K/AKT/mTOR signaling in cancer
  • Life Sciences › Biochemistry, Genetics and Molecular Biology › Molecular Biology › Protein Kinase Regulation and GTPase Signaling
  • Life Sciences › Biochemistry, Genetics and Molecular Biology › Molecular Biology › Metabolism, Diabetes, and Cancer
Keywords
PIK3CA, PIK3R1, p110α, p85α, oncogenic mutations, lipid binding, kinase activation mechanism, SH2 domain, activation loop, hydrophobic motif, allosteric regulation, conformational change, PIK3CA H1047R
MeSH terms
Phosphoinositide-3 Kinase Inhibitors, Amino Acid Sequence, Aniline Compounds, Animals, Cholesterol, Chromones, Enzyme Activation, Humans, Liposomes, Models, Molecular, Molecular Sequence Data, Phosphatidylcholines, Phosphatidylethanolamines, Phosphatidylserines, Phosphopeptides, Protein Binding, Sequence Deletion, Protein Structure, Secondary, Crystallography, X-Ray, Amino Acid Substitution, Catalytic Domain, Enzyme Activators, Protein Structure, Quaternary, Mice, Hydrophobic and Hydrophilic Interactions, Class I Phosphatidylinositol 3-Kinases, Class Ia Phosphatidylinositol 3-Kinase

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