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. 2009 Sep;10(9):990-6.
doi: 10.1038/embor.2009.126. Epub 2009 Jul 17.

Structural basis for recruitment of BRCA2 by PALB2

Antony W Oliver  1 , Sally Swift, Christopher J Lord, Alan Ashworth, Laurence H Pearl
Affiliations

Structural basis for recruitment of BRCA2 by PALB2

Antony W Oliver et al. EMBO Rep. 2009 Sep.

Erratum in

Abstract

The breast cancer 2, early onset protein (BRCA2) is central to the repair of DNA damage by homologous recombination. BRCA2 recruits the recombinase RAD51 to sites of damage, regulates its assembly into nucleoprotein filaments and thereby promotes homologous recombination. Localization of BRCA2 to nuclear foci requires its association with the partner and localizer of BRCA2 (PALB2), mutations in which are associated with cancer predisposition, as well as subtype N of Fanconi anaemia. We have determined the structure of the PALB2 carboxy-terminal beta-propeller domain in complex with a BRCA2 peptide. The structure shows the molecular determinants of this important protein-protein interaction and explains the effects of both cancer-associated truncating mutants in PALB2 and missense mutations in the amino-terminal region of BRCA2.

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Conflict of interest statement

The authors declare that they have no conflict of interest.

Figures

Figure 1
Figure 1
Structure of the PALB2 carboxy-terminal domain. (A) Bioinformatics analysis identifies a WD40-repeat domain forming the C-terminal part of PALB2 whereas the N-terminal and central regions are predicted to contain a coiled-coil structure. The boxed region indicates the amino-acid boundaries of the PALB2-C expression construct used in this study. (B) PALB2-C is a seven-bladed β-propeller (rainbow coloured blue → red, N → C terminus). Blade 7 is formed by strands A, B and C from the C-terminus and strand D from the N-terminus, which seal the toriodal structure in a ‘molecular Velcro' interaction. (C) Details of strand C of blade 7 and its environment. In families with Fanconi anaemia that have Y1183X mutations, the absence of C-terminal residues (magenta) prevents the closure of the WD40 ring and destabilizes the PALB2 protein, which is undetectable in these patients. PALB2, partner and localizer of BRCA2.
Figure 2
Figure 2
A minimal PALB2-binding motif in the BRCA2 amino terminus. (A) The previously localized PALB2-binding site in residues 10–40 of BRCA2 (Xia et al, 2006) was mapped to residues 21–39, encapsulating a minimal ten-residue motif, which is structured in the PALB2-C–BRCA2(21–39) complex. (B) Biotinylated wild-type BRCA2(21–39) peptide precipitates the PALB2-C construct, confirming it as the BRCA2-binding site. A peptide containing a breast-cancer-associated W31C mutation failed to co-precipitate PALB2-C. Bands marked with an asterisk arise from the NeutrAvidin resin. (C) Fluorescence polarization binding isotherm for interaction of PALB2-C with fluorescein-labelled BRCA2(21–39) peptide (see Methods). Kd as determined by nonlinear fitting of the data with a one-site-specific binding model (GraphPad Prism 5.0). BRCA2, breast cancer 2, early onset; PALB2, partner and localizer of BRCA2.
Figure 3
Figure 3
Structure of PALB2-C–BRCA2(21–39) complex. (A) BRCA2(21–39) (magenta) binds across a hydrophobic pocket at the crossover between blades 4 and 5 of the PALB2-C β-propeller, forming a short α-helix. The r.m.s.d. between the apo-bound and peptide-bound PALB2-C structures is 0.329 Å. (B) Molecular surface of PALB2-C (grey) highlighting the location of BRCA2 binding (magenta) compared with other β-propeller–peptide complexes. The superimposed eh1 motif from a complex with the WD40 domain of TLE1 (Jennings et al, 2006) marks the ‘classic' peptide motif-binding site found in most other WD40–peptide complexes. The positions of β-arrestin 2 and histone H4 peptides (blue), from complexes with the WD40 domains of clathrin and RBBP7, respectively (ter Haar et al, 2000; Murzina et al, 2008), are known examples of non-canonical sites, but are distinct from that observed for the PALB2-C–BRCA2(21–39) complex. (C) Details of the PALB2-C–BRCA2(21–39) interface. The core of the interaction is provided by Trp 31, Phe 32 and Leu 35 inserting into a hydrophobic pocket on PALB2-C (see text). The complex buries ∼1,100 Å2 of the molecular surface, typical of a reversible regulatory interaction. (D) Immunoprecipitation of FLAG–HA-tagged wild-type PALB2 and PALB2 with missense mutations in residues contributing to the BRCA2-binding site (see Methods). Endogenous BRCA2 is precipitated by wild-type PALB2, and to a lesser degree by the Met 1022 mutants. However, BRCA2 binding is abolished by mutation of Ala 1025, which lies at the bottom of the PALB2-hydrophobic pocket. The light chain of the M2 FLAG antibody provides a loading control. BRCA2, breast cancer 2, early onset; HA, haemagglutinin; PALB2, partner and localizer of BRCA2; TLE1, transducin-like enhancer of split 1.
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References

    1. Daniels MJ, Wang Y, Lee M, Venkitaraman AR (2004) Abnormal cytokinesis in cells deficient in the breast cancer susceptibility protein BRCA2. Science 306: 876–879 - PubMed
    1. Davies OR, Pellegrini L (2007) Interaction with the BRCA2 C terminus protects RAD51-DNA filaments from disassembly by BRC repeats. Nat Struct Mol Biol 14: 475–483 - PMC - PubMed
    1. Edwards SL, Brough R, Lord CJ, Natrajan R, Vatcheva R, Levine DA, Boyd J, Reis-Filho JS, Ashworth A (2008) Resistance to therapy caused by intragenic deletion in BRCA2. Nature 451: 1111–1115 - PubMed
    1. Esashi F, Galkin VE, Yu X, Egelman EH, West SC (2007) Stabilization of RAD51 nucleoprotein filaments by the C-terminal region of BRCA2. Nat Struct Mol Biol 14: 468–474 - PubMed
    1. Fuks F, Milner J, Kouzarides T (1998) BRCA2 associates with acetyltransferase activity when bound to P/CAF. Oncogene 17: 2531–2534 - PubMed

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