Structural insights into phosphoinositide 3-kinase catalysis and signalling

Phosphoinositide 3-kinases (PI3Ks) are ubiquitous lipid kinases that function both as signal transducers downstream of cell-surface receptors and in constitutive intracellular membrane and protein trafficking pathways. All PI3Ks are dual-specificity enzymes with a lipid kinase activity which phosphorylates phosphoinositides at the 3-hydroxyl, and a protein kinase activity. The products of PI3K-catalysed reactions, phosphatidylinositol 3,4,5-trisphosphate (PtdIns(3,4,5)P3), PtdIns(3,4)P2 and PtdIns(3)P, are second messengers in a variety of signal transduction pathways, including those essential to cell proliferation, adhesion, survival, cytoskeletal rearrangement and vesicle trafficking. Here we report the 2.2 Å X-ray crystallographic structure of the catalytic subunit of PI3Kγ, the class I enzyme that is activated by heterotrimeric G-protein βγ subunits and Ras. PI3Kγ has a modular organization centred around a helical-domain spine, with C2 and catalytic domains positioned to interact with phospholipid membranes, and a Ras-binding domain placed against the catalytic domain where it could drive allosteric activation of the enzyme.

[1]  J. Barrett,et al.  Differential regulation of p21 by p53 and Rb in cellular response to oxidative stress , 1999, Molecular carcinogenesis.

[2]  Michio Tsuda,et al.  A mutation in succinate dehydrogenase cytochrome b causes oxidative stress and ageing in nematodes , 1998, Nature.

[3]  Lan Huang,et al.  Structural basis for the interaction of Ras with RaIGDS , 1998, Nature Structural Biology.

[4]  L. Pirola,et al.  Bifurcation of lipid and protein kinase signals of PI3Kgamma to the protein kinases PKB and MAPK. , 1998, Science.

[5]  P. Cole,et al.  MEASUREMENT OF A BRONSTED NUCLEOPHILE COEFFICIENT AND INSIGHTS INTO THE TRANSITION STATE FOR A PROTEIN TYROSINE KINASE , 1997 .

[6]  P. Kraulis.  A program to produce both detailed and schematic plots of protein structures , 1991 .

[7]  G. Thomas,et al.  Target of rapamycin (TOR): balancing the opposing forces of protein synthesis and degradation. , 1999, Current opinion in genetics & development.

[8]  A. Wolfman,et al.  Different structural requirements within the switch II region of the Ras protein for interactions with specific downstream targets. , 1995, Oncogene.

[9]  S. Taylor,et al.  Catalytic subunit of cyclic AMP-dependent protein kinase: structure and dynamics of the active site cleft. , 1999, Pharmacology & therapeutics.

[10]  T. Johnson,et al.  A genetic pathway conferring life extension and resistance to UV stress in Caenorhabditis elegans. , 1996, Genetics.

[11]  R. Weindruch,et al.  Dietary restriction in mice beginning at 1 year of age: effect on life-span and spontaneous cancer incidence. , 1982, Science.

[12]  Alfred Wittinghofer,et al.  Structural View of the Ran–Importin β Interaction at 2.3 Å Resolution , 1999, Cell.

[13]  S. Benzer,et al.  Extended life-span and stress resistance in the Drosophila mutant methuselah. , 1998, Science.

[14]  M. Chalfie,et al.  A cytosolic catalase is needed to extend adult lifespan in C. elegans daf-C and clk-1 mutants , 1999, Nature.

[15]  S. Emr,et al.  Vps34p required for yeast vacuolar protein sorting is a multiple specificity kinase that exhibits both protein kinase and phosphatidylinositol-specific PI 3-kinase activities. , 1994, The Journal of biological chemistry.

[16]  Lan Huang,et al.  Three-dimensional structure of the Ras-interacting domain of RalGDS , 1997, Nature Structural Biology.

[17]  M. Zvelebil,et al.  Wortmannin inactivates phosphoinositide 3-kinase by covalent modification of Lys-802, a residue involved in the phosphate transfer reaction , 1996, Molecular and cellular biology.

[18]  B. Kennedy,et al.  Mutation in the silencing gene S/R4 can delay aging in S. cerevisiae , 1995, Cell.

[19]  M. Waterfield,et al.  Using structure to define the function of phosphoinositide 3‐kinase family members , 1997, FEBS letters.

[20]  Roger L. Williams,et al.  A ternary metal binding site in the C2 domain of phosphoinositide-specific phospholipase C-delta1. , 1997, Biochemistry.

[21]  P. Warne,et al.  Role of Phosphoinositide 3-OH Kinase in Cell Transformation and Control of the Actin Cytoskeleton by Ras , 1997, Cell.

[22]  E. Nånberg,et al.  Role of phosphoinositide 3OH‐kinase in autocrine transformation by PDGF‐BB * , 2001, Journal of cellular physiology.

[23]  P Griffiths,et al.  Mice with a Homozygous Null Mutation for the Most Abundant Glutathione Peroxidase, Gpx1, Show Increased Susceptibility to the Oxidative Stress-inducing Agents Paraquat and Hydrogen Peroxide* , 1998, The Journal of Biological Chemistry.

[24]  R. Weindruch,et al.  Oxidative Stress, Caloric Restriction, and Aging , 1996, Science.

[25]  P. Hawkins,et al.  The Gβγ Sensitivity of a PI3K Is Dependent upon a Tightly Associated Adaptor, p101 , 1997, Cell.

[26]  S. Volinia,et al.  Cloning and characterization of a G protein-activated human phosphoinositide-3 kinase. , 1995, Science.

[27]  R. Weindruch,et al.  The retardation of aging in mice by dietary restriction: longevity, cancer, immunity and lifetime energy intake. , 1986, The Journal of nutrition.

[28]  Brian A. Hemmings,et al.  The Structure of the Protein Phosphatase 2A PR65/A Subunit Reveals the Conformation of Its 15 Tandemly Repeated HEAT Motifs , 1999, Cell.

[29]  Thomas C. Terwilliger,et al.  Automated MAD and MIR structure solution , 1999, Acta crystallographica. Section D, Biological crystallography.

[30]  Collaborative Computational,et al.  The CCP4 suite: programs for protein crystallography. , 1994, Acta crystallographica. Section D, Biological crystallography.

[31]  G. Bricogne,et al.  [27] Maximum-likelihood heavy-atom parameter refinement for multiple isomorphous replacement and multiwavelength anomalous diffraction methods. , 1997, Methods in enzymology.

[32]  I Gout,et al.  PI 3‐kinase is a dual specificity enzyme: autoregulation by an intrinsic protein‐serine kinase activity. , 1994, The EMBO journal.

[33]  P. Hawkins,et al.  Characterizing the Interactions between the Two Subunits of the p101/p110γ Phosphoinositide 3-Kinase and Their Role in the Activation of This Enzyme by Gβγ Subunits* , 1999, The Journal of Biological Chemistry.

[34]  J. Zou,et al.  Improved methods for building protein models in electron density maps and the location of errors in these models. , 1991, Acta crystallographica. Section A, Foundations of crystallography.

[35]  E. W. Hutchinson,et al.  Resistance to Environmental Stress in Drosophila melanogaster Selected for Postponed Senescence , 1985, Physiological Zoology.

[36]  A. Wittinghofer,et al.  The 2.2 Å crystal structure of the Ras-binding domain of the serine/threonine kinase c-Raf1 in complex with RaplA and a GTP analogue , 1995, Nature.

[37]  T. Kirkwood,et al.  Mechanisms and Evolution of Aging , 1996, Science.

[38]  Hiroto Yamaguchi,et al.  Structural basis for activation of human lymphocyte kinase Lck upon tyrosine phosphorylation , 1996, Nature.

[39]  R. S. Sohal,et al.  Extension of life-span by overexpression of superoxide dismutase and catalase in Drosophila melanogaster. , 1994, Science.

[40]  K. Sharp,et al.  Protein folding and association: Insights from the interfacial and thermodynamic properties of hydrocarbons , 1991, Proteins.

[41]  C. Marshall.  Ras effectors. , 1996, Current opinion in cell biology.

[42]  A. Toker,et al.  Signalling through the lipid products of phosphoinositide-3-OH kinase , 1997, Nature.

[43]  J. Hurley,et al.  Structure of Type IIβ Phosphatidylinositol Phosphate Kinase A Protein Kinase Fold Flattened for Interfacial Phosphorylation , 1998, Cell.

[44]  D. Riddle,et al.  Genes that regulate both development and longevity in Caenorhabditis elegans. , 1995, Genetics.

[45]  R J Read,et al.  Crystallography & NMR system: A new software suite for macromolecular structure determination. , 1998, Acta crystallographica. Section D, Biological crystallography.

[46]  M. Waterfield,et al.  Autophosphorylation of p110δ phosphoinositide 3‐kinase: a new paradigm for the regulation of lipid kinases in vitro and in vivo , 1999, The EMBO journal.

[47]  P. Medawar.  Old Age and Natural Death , 2019, The Uniqueness of the Individual.