Structural basis for recognition of H3T3ph and Smac/DIABLO N-terminal peptides by human Survivin - PubMed (original) (raw)

Structural basis for recognition of H3T3ph and Smac/DIABLO N-terminal peptides by human Survivin

Jiamu Du et al. Structure. 2012.

Abstract

Survivin is an inhibitor of apoptosis family protein implicated in apoptosis and mitosis. In apoptosis, it has been shown to recognize the Smac/DIABLO protein. It is also a component of the chromosomal passenger complex, a key player during mitosis. Recently, Survivin was identified in vitro and in vivo as the direct binding partner for phosphorylated Thr3 on histone H3 (H3T3ph). We have undertaken structural and binding studies to investigate the molecular basis underlying recognition of H3T3ph and Smac/DIABLO N-terminal peptides by Survivin. Our crystallographic studies establish recognition of N-terminal Ala in both complexes and identify intermolecular hydrogen-bonding interactions in the Survivin phosphate-binding pocket that contribute to H3T3ph mark recognition. In addition, our calorimetric data establish that Survivin binds tighter to the H3T3ph-containing peptide relative to the N-terminal Smac/DIABLO peptide, and this preference can be reversed through structure-guided mutations that increase the hydrophobicity of the phosphate-binding pocket.

Copyright © 2012 Elsevier Ltd. All rights reserved.

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Figures

Figure 1

Figure 1. Interactions between H3(1–15)T3ph Peptide and Survivin in the Complex

  1. Overall interaction between residues Ala1 to Gln5 of H3(1–15)T3ph peptide and Survivin in the complex. The bound peptide (yellow) in a stick representation is positioned within the BIR domain (cyan) of Survivin in a ribbon representation.
  2. An electrostatics surface representation of Survivin with bound residues Ala1 to Gln5 of H3(1–15)T3ph peptide (yellow) in a stick representation. The N-terminus and Ala1 insert into a negatively charged pocket, the Arg2 and Lys4 side chains lie on a flat negatively charge surface, while Thr3ph lies on a positively charged shallow cleft. There is electrostatic complementarity between the bound H3T3ph peptide and Survivin in the complex.
  3. Stereo view highlighting details of the intermolecular interactions between the A1-R2-T3ph-K4 segment of the bound H3(1–15)T3ph peptide and binding pocket residues of the BIR domain of Survivin. Intermolecular hydrogen-bonding interactions are designated by dashed red lines.
  4. Stereo view of superpositioned structures of unliganded Survivin (grey), H3(1–15)T3ph peptide-bound Survivin (cyan) and H3(1–10) peptide-bound Survivin (magenta). These views emphasize binding pocket interactions. Note the shift in the α-helix (on left) on complex formation. See also Supplementary Figures S1, S2, S4, S5, S6 and S7.

Figure 2

Figure 2. Interactions between SmacN(1–15) Peptide and Survivin in the Complex

  1. Overall interaction between residues Ala1 to Ile4 of SmacN(1–15) peptide and Survivin in the complex. The bound peptide (yellow) in a stick representation is positioned within the BIR domain (cyan) of Survivin in a ribbon representation.
  2. An electrostatics surface representation of Survivin with bound residues Ala1 to Ile4 of SmacN peptide (yellow) in a stick representation.
  3. Stereo view highlighting details of the intermolecular interactions between the A1-V2-P3-I4 segment of the bound SmacN(1–15) peptide and binding pocket residues of the BIR domain of Survivin. Intermolecular hydrogen-bonding interactions are designated by dashed red lines.
  4. Stereo view of superpositioned structures of unliganded Survivin (grey) and SmacN peptide-bound Survivin (cyan). These views emphasize binding pocket interactions. Note the shift in the α-helix (on left) on complex formation.
    See also Supplementary Figures S2 and S7.

Figure 3

Figure 3. Pull-down and Isothermal Titration Calorimetry (ITC) Measurements of Binding Affinities between Survivin and H3 Peptides containing Modifications

  1. A pull-down assay testing binding of human Survivin to histone H3 containing different modifications. Purified human Survivin (1–142) was incubated with the indicated peptide beads. Coomassie staining of input and bead fractions is shown.
  2. ITC binding curve for complex formation between H3(1–15)T3ph peptide and Survivin.
  3. ITC binding curve for complex formation between H3(1–10) peptide and Survivin.
  4. ITC binding curve for complex formation between H3(1–20)R2me2aT3ph peptide and Survivin.
  5. ITC binding curve for complex formation between H3(1–20)T3phK4me3 peptide and Survivin.

Figure 4

Figure 4. Interactions between SmacN(1–15) Peptide and Survivin (K62Y/H80W) Mutants in the Complex

  1. ITC binding curves for complex formation between SmacN(1–15) peptide and wild-type Survivin (red curve) and Survivin (K62Y/H80W) mutant (blue curve).
  2. An electrostatics surface representation of Survivin (K62R/H80W) mutant with bound residues A1 to I4 of SmacN(1–15) peptide (yellow) in a stick representation. The N-terminus and Ala1 insert into a negatively charged pocket, while the Pro3 ring lies on a hydrophobic surface formed by Tyr62 and Trp80.
  3. Stereo view highlighting details of the intermolecular interactions between the A1-V2-P3-I4 segment of the bound SmacN(1–15) peptide and binding pocket residues of the BIR domain of Survivin (K62R/H80W) mutant. Intermolecular hydrogen-bonding interactions are designated by dashed red lines.
    See also Supplementary Figure 3.

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