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Yersinia effector YopO uses actin as bait to phosphorylate proteins that regulate actin polymerization

Nature Structural & Molecular Biology volume 22, pages 248–255 (2015)Cite this article

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Abstract

Pathogenic Yersinia species evade host immune systems through the injection of Yersinia outer proteins (Yops) into phagocytic cells. One Yop, YopO, also known as YpkA, induces actin-filament disruption, impairing phagocytosis. Here we describe the X-ray structure of Yersinia enterocolitica YopO in complex with actin, which reveals that YopO binds to an actin monomer in a manner that blocks polymerization yet allows the bound actin to interact with host actin-regulating proteins. SILAC-MS and biochemical analyses confirm that actin-polymerization regulators such as VASP, EVL, WASP, gelsolin and the formin diaphanous 1 are directly sequestered and phosphorylated by YopO through formation of ternary complexes with actin. This leads to a model in which YopO at the membrane sequesters actin from polymerization while using the bound actin as bait to recruit, phosphorylate and misregulate host actin-regulating proteins to disrupt phagocytosis.

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Figure 1: Structure of the YopO–actin complex.
Figure 2: The YopO–actin complex is polymerization incompetent.
Figure 3: Structural basis of differential actin isoform preference of YopO.
Figure 4: YopO-bound actin interacts with other actin-binding proteins.
Figure 5: Phosphorylation of actin-binding proteins by YopO.
Figure 6: Model for disabling actin polymerization by YopO.

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References

  1. Aderem, A. & Underhill, D. Mechanisms of phagocytosis in macrophages. Annu. Rev. Immunol. 17, 593–623 (1999).

    Article  CAS  Google Scholar 

  2. Griffin, F.M., Griffin, J.A., Leider, J.E. & Silverstein, S.C. Studies on the mechanism of phagocytosis: requirements for circumferential attachment of particle-bound ligands to specific receptors on the macrophage plasma membrane. J. Exp. Med. 142, 1263–1282 (1975).

    Article  Google Scholar 

  3. Witke, W., Li, W., Kwiatkowski, D.J. & Southwick, F.S. Comparisons of CapG and gelsolin-null macrophages: demonstration of a unique role for CapG in receptor-mediated ruffling, phagocytosis, and vesicle rocketing. J. Cell Biol. 154, 775–784 (2001).

    Article  CAS  Google Scholar 

  4. Serrander, L. et al. Selective inhibition of IgG-mediated phagocytosis in gelsolin-deficient murine neutrophils. J. Immunol. 165, 2451–2457 (2000).

    Article  CAS  Google Scholar 

  5. Colucci-Guyon, E. et al. A role for mammalian diaphanous-related formins in complement receptor (CR3)-mediated phagocytosis in macrophages. Curr. Biol. 15, 2007–2012 (2005).

    Article  CAS  Google Scholar 

  6. Lorenzi, R., Brickell, P.M., Katz, D.R., Kinnon, C. & Thrasher, A.J. Wiskott-Aldrich syndrome protein is necessary for efficient IgG-mediated phagocytosis. Blood 95, 2943–2946 (2000).

    CAS  PubMed  Google Scholar 

  7. Tsuboi, S. & Meerloo, J. Wiskott-Aldrich syndrome protein is a key regulator of the phagocytic cup formation in macrophages. J. Biol. Chem. 282, 34194–34203 (2007).

    Article  CAS  Google Scholar 

  8. Coppolino, M.G. et al. Evidence for a molecular complex consisting of Fyb/SLAP, SLP-76, Nck, VASP and WASP that links the actin cytoskeleton to Fcγ receptor signalling during phagocytosis. J. Cell Sci. 114, 4307–4318 (2001).

    CAS  PubMed  Google Scholar 

  9. Cornelis, G.R. Molecular and cell biology aspects of plague. Proc. Natl. Acad. Sci. USA 97, 8778–8783 (2000).

    Article  CAS  Google Scholar 

  10. Harbeck, M. et al. Yersinia pestis DNA from skeletal remains from the 6th century AD reveals insights into Justinianic Plague. PLoS Pathog. 9, e1003349 (2013).

    Article  CAS  Google Scholar 

  11. Haensch, S. et al. Distinct clones of Yersinia pestis caused the black death. PLoS Pathog. 6, e1001134 (2010).

    Article  Google Scholar 

  12. Perry, R.D. & Fetherston, J.D. Yersinia pestis: etiologic agent of plague. Clin. Microbiol. Rev. 10, 35–66 (1997).

    Article  CAS  Google Scholar 

  13. Butler, T. Plague gives surprises in the first decade of the 21st century in the United States and worldwide. Am. J. Trop. Med. Hyg. 89, 788–793 (2013).

    Article  Google Scholar 

  14. Galimand, M., Carniel, E. & Courvalin, P. Resistance of Yersinia pestis to antimicrobial agents. Antimicrob. Agents Chemother. 50, 3233–3236 (2006).

    Article  CAS  Google Scholar 

  15. Trasak, C. et al. Yersinia protein kinase YopO is activated by a novel G-actin binding process. J. Biol. Chem. 282, 2268–2277 (2007).

    Article  CAS  Google Scholar 

  16. Letzelter, M. et al. The discovery of SycO highlights a new function for type III secretion effector chaperones. EMBO J. 25, 3223–3233 (2006).

    Article  CAS  Google Scholar 

  17. Håkansson, S., Galyov, E.E., Rosqvist, R. & Wolf-Watz, H. The Yersinia YpkA Ser/Thr kinase is translocated and subsequently targeted to the inner surface of the HeLa cell plasma membrane. Mol. Microbiol. 20, 593–603 (1996).

    Article  Google Scholar 

  18. Galyov, E.E., Håkansson, S., Forsberg, A. & Wolf-Watz, H. A secreted protein kinase of Yersinia pseudotuberculosis is an indispensable virulence determinant. Nature 361, 730–732 (1993).

    Article  CAS  Google Scholar 

  19. Juris, S.J., Rudolph, A.E., Huddler, D., Orth, K. & Dixon, J.E. A distinctive role for the Yersinia protein kinase: actin binding, kinase activation, and cytoskeleton disruption. Proc. Natl. Acad. Sci. USA 97, 9431–9436 (2000).

    Article  CAS  Google Scholar 

  20. Prehna, G., Ivanov, M.I., Bliska, J.B. & Stebbins, C.E. Yersinia virulence depends on mimicry of host Rho-family nucleotide dissociation inhibitors. Cell 126, 869–880 (2006).

    Article  CAS  Google Scholar 

  21. Dukuzumuremyi, J.M. et al. The Yersinia protein kinase A is a host factor inducible RhoA/Rac-binding virulence factor. J. Biol. Chem. 275, 35281–35290 (2000).

    Article  CAS  Google Scholar 

  22. Wiley, D.J. et al. The Ser/Thr kinase activity of the Yersinia protein kinase A (YpkA) is necessary for full virulence in the mouse, mollifying phagocytes, and disrupting the eukaryotic cytoskeleton. Microb. Pathog. 40, 234–243 (2006).

    Article  CAS  Google Scholar 

  23. Navarro, L. et al. Identification of a molecular target for the Yersinia protein kinase A. Mol. Cell 26, 465–477 (2007).

    Article  CAS  Google Scholar 

  24. Cooper, D.R. et al. Protein crystallization by surface entropy reduction: optimization of the SER strategy. Acta Crystallogr. D Biol. Crystallogr. 63, 636–645 (2007).

    Article  CAS  Google Scholar 

  25. Wang, H., Robinson, R.C. & Burtnick, L.D. The structure of native G-actin. Cytoskeleton (Hoboken) 67, 456–465 (2010).

    Article  CAS  Google Scholar 

  26. Fujii, T., Iwane, A.H., Yanagida, T. & Namba, K. Direct visualization of secondary structures of F-actin by electron cryomicroscopy. Nature 467, 724–728 (2010).

    Article  CAS  Google Scholar 

  27. Dominguez, R. Actin-binding proteins: a unifying hypothesis. Trends Biochem. Sci. 29, 572–578 (2004).

    Article  CAS  Google Scholar 

  28. Xue, B. & Robinson, R.C. Guardians of the actin monomer. Eur. J. Cell Biol. 92, 316–332 (2013).

    Article  CAS  Google Scholar 

  29. Ong, S.-E. & Mann, M. A practical recipe for stable isotope labeling by amino acids in cell culture (SILAC). Nat. Protoc. 1, 2650–2660 (2006).

    Article  CAS  Google Scholar 

  30. Ferron, F., Rebowski, G., Lee, S.H. & Dominguez, R. Structural basis for the recruitment of profilin-actin complexes during filament elongation by Ena/VASP. EMBO J. 26, 4597–4606 (2007).

    Article  CAS  Google Scholar 

  31. Burtnick, L.D., Urosev, D., Irobi, E., Narayan, K. & Robinson, R.C. Structure of the N-terminal half of gelsolin bound to actin: roles in severing, apoptosis and FAF. EMBO J. 23, 2713–2722 (2004).

    Article  CAS  Google Scholar 

  32. Chereau, D. et al. Actin-bound structures of Wiskott–Aldrich syndrome protein (WASP)-homology domain 2 and the implications for filament assembly. Proc. Natl. Acad. Sci. USA 102, 16644–16649 (2005).

    Article  CAS  Google Scholar 

  33. Lee, S.H. et al. Structural basis for the actin-binding function of missing-in-metastasis. Structure 15, 145–155 (2007).

    Article  Google Scholar 

  34. Bryan, J. Gelsolin has three actin-binding sites. J. Cell Biol. 106, 1553–1562 (1988).

    Article  CAS  Google Scholar 

  35. Gieselmann, R., Kwiatkowski, D.J., Janmey, P.A. & Witke, W. Distinct biochemical characteristics of the two human profilin isoforms. Eur. J. Biochem. 229, 621–628 (1995).

    Article  CAS  Google Scholar 

  36. Trülzsch, K., Sporleder, T., Igwe, E.I., Russmann, H. & Heesemann, J. Contribution of the major secreted yops of Yersinia enterocolitica O:8 to pathogenicity in the mouse infection model. Infect. Immun. 72, 5227–5234 (2004).

    Article  Google Scholar 

  37. Delorme-Walker, V. et al. Toxofilin upregulates the host cortical actin cytoskeleton dynamics, facilitating Toxoplasma invasion. J. Cell Sci. 125, 4333–4342 (2012).

    Article  CAS  Google Scholar 

  38. Aktories, K. Bacterial protein toxins that modify host regulatory GTPases. Nat. Rev. Microbiol. 9, 487–498 (2011).

    Article  CAS  Google Scholar 

  39. Calle, Y., Anton, I., Thrasher, A.J. & Jones, G.E. WASP and WIP regulate podosomes in migrating leukocytes. J. Microsc. 231, 494–505 (2008).

    Article  CAS  Google Scholar 

  40. Döppler, H. & Storz, P. Regulation of VASP by phosphorylation. Cell Adh. Migr. 7, 482–486 (2013).

    Article  Google Scholar 

  41. Nag, S., Larsson, M., Robinson, R.C. & Burtnick, L.D. Gelsolin: the tail of a molecular gymnast. Cytoskeleton (Hoboken) 70, 360–384 (2013).

    Article  CAS  Google Scholar 

  42. Li, D., Dammer, E.B., Lucki, N.C. & Sewer, M.B. cAMP-stimulated phosphorylation of diaphanous 1 regulates protein stability and interaction with binding partners in adrenocortical cells. Mol. Biol. Cell 24, 848–857 (2013).

    Article  Google Scholar 

  43. Groves, E. et al. Sequestering of Rac by the Yersinia effector YopO blocks Fc receptor-mediated phagocytosis. J. Biol. Chem. 285, 4087–4098 (2010).

    Article  CAS  Google Scholar 

  44. Park, S.K., Venable, J.D., Xu, T. & Yates, J.R. III. A quantitative analysis software tool for mass spectrometry–based proteomics. Nat. Methods 5, 319–322 (2008).

    Article  CAS  Google Scholar 

  45. Ohki, T., Ohno, C., Oyama, K., Mikhailenko, S.V. & Ishiwata, S. Purification of cytoplasmic actin by affinity chromatography using the C-terminal half of gelsolin. Biochem. Biophys. Res. Commun. 383, 146–150 (2009).

    Article  CAS  Google Scholar 

  46. Goldschmidt, L., Cooper, D.R., Derewenda, Z.S. & Eisenberg, D. Toward rational protein crystallization: a Web server for the design of crystallizable protein variants. Protein Sci. 16, 1569–1576 (2007).

    Article  CAS  Google Scholar 

  47. Kabsch, W. Automatic processing of rotation diffraction data from crystals of initially unknown symmetry and cell constants. J. Appl. Crystallogr. 26, 795–800 (1993).

    Article  CAS  Google Scholar 

  48. Winter, G., Lobley, C.M.C. & Prince, S.M. Decision making in xia2. Acta Crystallogr. D Biol. Crystallogr. 69, 1260–1273 (2013).

    Article  CAS  Google Scholar 

  49. Evans, P.R. & Murshudov, G.N. How good are my data and what is the resolution? Acta Crystallogr. D Biol. Crystallogr. 69, 1204–1214 (2013).

    Article  CAS  Google Scholar 

  50. Otwinowski, Z. & Minor, W. Processing of X-ray diffraction data collected in oscillation mode. Methods Enzymol. 276, 307–326 (1997).

    Article  CAS  Google Scholar 

  51. Evans, P. Scaling and assessment of data quality. Acta Crystallogr. D Biol. Crystallogr. 62, 72–82 (2006).

    Article  Google Scholar 

  52. Zwart, P.H., Afonine, P.V. & Grosse-Kunstleve, R.W. Automated structure solution with the PHENIX suite. Methods Mol. Biol. 426, 419–435 (2008).

    Article  CAS  Google Scholar 

  53. Malakhova, M. et al. The crystal structure of the active form of the C-terminal kinase domain of mitogen- and stress-activated protein kinase 1. J. Mol. Biol. 399, 41–52 (2010).

    Article  CAS  Google Scholar 

  54. Stein, N. CHAINSAW: a program for mutating pdb files used as templates in molecular replacement. J. Appl. Crystallogr. 41, 641–643 (2008).

    Article  CAS  Google Scholar 

  55. Cowtan, K. Recent developments in classical density modification. Acta Crystallogr. D Biol. Crystallogr. 66, 470–478 (2010).

    Article  CAS  Google Scholar 

  56. DiMaio, F. et al. Improved molecular replacement by density- and energy-guided protein structure optimization. Nature 473, 540–543 (2011).

    Article  CAS  Google Scholar 

  57. Emsley, P., Lohkamp, B., Scott, W.G. & Cowtan, K. Features and development of Coot. Acta Crystallogr. D Biol. Crystallogr. 66, 486–501 (2010).

    Article  CAS  Google Scholar 

  58. Terwilliger, T.C. et al. Iterative-build OMIT maps: map improvement by iterative model building and refinement without model bias. Acta Crystallogr. D Biol. Crystallogr. 64, 515–524 (2008).

    Article  CAS  Google Scholar 

  59. Xu, T. et al. ProLuCID, a fast and sensitive tandem mass spectra-based protein identification program. Mol. Cell. Proteomics 5, S174 (2006).

    Google Scholar 

  60. Tabb, D.L., Mcdonald, W.H. & Yates, J.R. DTASelect and Contrast: tools for assembling and comparing protein identifications from shotgun proteomics. J. Proteome Res. 1, 21–26 (2002).

    Article  CAS  Google Scholar 

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Acknowledgements

We thank L. Burtnick and W. Burkholder for discussions and critical reading of the manuscript. W.L.L. and R.C.R. thank the A*STAR for support. We acknowledge the Joint Centre for Structural Biology, Singapore, which is supported by Nanyang Technological University and the Biomedical Research Council (BMRC) of A*STAR, for providing research facilities and P. Kaldis for providing help and facilities for the kinase assays. We thank the Diamond Light Source (proposal MX8423) for crystal screening and beamline BL13B1 at the National Synchrotron Radiation Research Center, Taiwan (NSRRC) for final data collection. The Wellcome Trust Centre for Human Genetics is supported by the Wellcome Trust Core award (090532/Z/09/Z). We thank L. Blanchoin (Institut de Recherches en Technologies et Sciences pour le Vivant) and M. Hernandez-Valladares (University of Liverpool) for reagents.

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Authors and Affiliations

  1. Institute of Molecular and Cell Biology, Agency for Science, Technology and Research (A*STAR), Singapore

    Wei Lin Lee & Robert C Robinson

  2. Division of Structural Biology, Wellcome Trust Centre for Human Genetics, University of Oxford, Oxford, UK

    Wei Lin Lee & Jonathan M Grimes

  3. Diamond Light Source, Oxfordshire, UK

    Jonathan M Grimes

  4. Department of Biochemistry, National University of Singapore, Singapore

    Robert C Robinson

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W.L.L. carried out the experimental work. All authors analyzed the data and prepared the manuscript.

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Integrated supplementary information

Supplementary Figure 1 The sequence and secondary structure of YopO and its interaction with actin.

Sequence/structural analysis of YopO of Yersinia enterocolitica (89-729) (accession NP_783721) aligned against YpkA of Yersinia pestis (accession NP_395206) and Yersinia pseudotuberculosis (accession YP_068414). The secondary structure of the kinase and GDI domains are indicated in blue and red above the sequence alignment, respectively. Disordered regions are represented by disconnected lines. The putative phosphorylation sites Ser90 and Ser95 are marked with black circles. The surface entropy reduction mutations are indicated by black asterisks. Residues that constitute the αC helix, the catalytic loop and the activation segment are shown in magenta, red and green, respectively. The catalytic residue Asp267 is marked with a red circle and residues involved in interaction with actin and Rac1 are indicated with cyan and yellow squares, respectively. The alignment was performed using Clustal Omega and drawn with ESPript.

Supplementary Figure 2 Details on the structure of the YopO–actin complex.

(a) Analysis of the surface entropy reduction mutations in the final structure. One of the mutations stabilizes a crystal contact. Actin is shown in cyan, the GDI domain in red and the kinase domain in blue, following the color scheme in Fig. 1a. One mutation on YopO (E207Y) interacts with a residue (Y588) on the GDI domain of a symmetry-related molecular complex, and is likely to be basis of the improvement in diffraction quality of the mutant protein crystals.

(b) Conformational changes to the GDI domain. The isolated GDI domain is shown in fawn (PDB: 2H7O), the Rac-bound GDI domain (PDB: 2H7V) is shown in light blue with Rac1 in yellow, and the GDI domain taken from the YopO:actin complex in red. The three GDI domains are superposed, as aligned on the N-terminal GDI subdomain.

(c) Comparison of the structural elements of the kinase domain of YopO (blue) with human PAK4 (PDB code 4FIJ, yellow) and human OSR1 (PDB code 2VWI, gray) (from left to right). The structural elements are colored following the scheme in Supplementary Note Figure 1. The P-loop is disordered in the YopO structure.

(d) Snapshots of the 2Fo-Fc (2σ, gray) experimental electron density omit map for different segments of the kinase domain.

Supplementary Figure 3 YopO mutants.

(a) Disrupting the YopO:actin interface in YopO nAct. YopO is colored as in Fig. 1a. Panels show close up of the interfaces between YopO and actin at the mutation sites, which were used to disrupt the interactions with actin in YopO nAct. Mutations V374D and T376D introduce large charged groups into a hydrophobic YopO:actin interface while mutations R723A and E727A disrupt electrostatic interactions between YopO and actin.

(b) Size exclusion chromatograms of YopO WT and mutants. YopO WT and mutants have a similar rate of migration on size exclusion chromatography, suggesting that the mutations do not disrupt the normal folding of YopO. WT, KD, nAct and nRac denote wild type, kinase-dead, non-actin binding and non-Rac binding respectively.

Supplementary Figure 4 Sequence alignment of actin protein sequences.

Sequences include S.frugiperda actin (SfACT, accession: AEK82131), D.discoideum actin (DdACT, AAA33145), human β-actin (hACTB, accession: NP_001092.1), mouse β-actin (mACTB, accession: AAA37164) and rabbit α-actin (aACTA, accession: CAA43139.1). The corresponding position for Thr202 of S.frugiperda is marked with a yellow box. Red and blue squares beneath the alignment represent interactions with the YopO GDI and kinase domains, respectively. The alignment was performed using Clustal Omega and presented using ESPript.

Supplementary Figure 5 YopO uses actin as bait for phosphorylation.

(a) Model of the ternary complex of YopO:actin:gelsolin (G4–G6), following the color scheme in Fig. 1a, with gelsolin in green (PDB code 1H1V). Note that gelsolin comprises six domains, two of which bind G-actin similarly, G1 and G4. CapG consists of three gelsolin domains. Models of YopO–actin–G1–3 (Fig. 4b) and YopO–actin–G4–6 represent potential models of the YopO–actin–CapG structure.

(b) Model of the ternary complex of YopO–actin–ADFH, with the ADFH of twinfilin in magenta (PDB code 3DAW). Note that this represents a model of the YopO–actin–cofilin structure. Full length twinfilin contains a second ADFH domain. CapG consists of three gelsolin domains and Twf1, of two ADF-H domains. Both CapG and Twf1 are not sufficiently elongated to span from the actin-binding site to the kinase catalytic cleft in order to be phosphorylated by YopO–actin.

(c) In vitro phosphorylation assay of YopO and mutants. YopO WT or mutants (4.7 μM) were incubated with or without Sf9 actin (4.7 μM), VASP as substrate (14.1 μM) or profilin (14.1 μM) as indicated, supplemented with 5μCi [γ-32P]ATP. Lane 1: YopO does not phosphorylate itself in the absence of actin. Lane 2: YopO autophosphorylates in the presence of actin. Lane 3: VASP is phosphorylated in the presence of actin and YopO; which is not impeded by the presence of profilin (lane 4). Lane 5: Kinase-dead (KD) YopO does not phosphorylate itself or VASP, and neither does nAct the reduced actin binding YopO mutant (lane 6). Lane 7: The non-Rac binding YopO mutant (nRac) phosphorylates itself and VASP.

(d) SDS-PAGE analysis of the proteins used in the in vitro phosphorylation assay in Fig. 5. YopO, VASP, EVL, gelsolin, CapG, twinfilin1 are without fusion tags. mDia1 (583–1262) and WASP (150–502) each contain both an N-terminal GST and a C-terminal His6 tag. Due to overlap in the molecular weights of YopO and GST-WASP, in Fig. 5, the phosphorylated GST-WASP was cleaved with thrombin prior to SDS-PAGE. In this figure, GST-WASP was not cleaved with thrombin. Red stars indicate the molecular weight of the substrates. Invitrogen Benchmark protein ladder is loaded in the first lane, with the molecular weights of bands in the ladder indicated. The gel was stained with Coomassie blue.

Supplementary Figure 6 Structural comparison of actin-binding proteins from pathogens and hypothetical models for structural activation of YopO.

(a) Superposition of toxofilin (PDB:2Q97) on the structure of YopO:actin, following the colour scheme in Fig. 1a, with toxofilin in yellow. The N- and C- terminal residues of the toxofilin construct are shown as yellow and magenta spheres, respectively. The C-terminus of toxofilin wraps around actin subdomain 4 in a manner similar to that of YopO, although the topologies and the exact interaction interfaces differ.

(b) Superposition of Iota toxin component Ia (PDB:3BUZ) on the structure of YopO:actin, following the colour scheme in Fig. 1a, with Iota toxin Ia in magenta.

(c) Hypothetical model of YopO activation. The GDI domain is shown in red and the kinase domain is shown in blue, with actin shown as a surface representation in marine. In this hypothetical model of the inactive structure, the backbone helix of the GDI domain would be straight and the kinase active site would likely be inaccessible. In the transition from the inactive YopO to YopO in the YopO:actin complex, two potential “intermediate” complexes are possible. Actin could first bind to either the kinase domain or the GDI domain, though the latter is more plausible due to a greater solvation free energy gain for the formation of this interface as compared to binding of actin to the kinase domain. The series of conformational changes that lead to sandwiching of actin between the two domains would lead to the bending of the backbone helix of the GDI domain and the kinase substrate binding groove being exposed as in the YopO:actin structure.

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Lee, W., Grimes, J. & Robinson, R. Yersinia effector YopO uses actin as bait to phosphorylate proteins that regulate actin polymerization. Nat Struct Mol Biol 22, 248–255 (2015). https://doi.org/10.1038/nsmb.2964

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