IQ motif selectivity in human IQGAP1: binding of myosin essential light chain and S100B

  • Sevvel Pathmanathan
  • Sarah F. Elliott
  • Sara McSwiggen
  • Brett Greer
  • Pat Harriott
  • G. Brent Irvine
  • David J. Timson
Article

Abstract

IQGAPs are cytoskeletal scaffolding proteins which link signalling pathways to the reorganisation of actin and microtubules. Human IQGAP1 has four IQ motifs each of which binds to calmodulin. The same region has been implicated in binding to two calmodulin-like proteins, the myosin essential light chain Mlc1sa and the calcium and zinc ion binding protein S100B. Using synthetic peptides corresponding to the four IQ motifs of human IQGAP1, we showed by native gel electrophoresis that only the first IQ motif interacts with Mlc1sa. This IQ motif, and also the fourth, interacts with the budding yeast myosin essential light chain Mlc1p. The first and second IQ motifs interact with S100B in the presence of calcium ions. This clearly establishes that S100B can interact with its targets through IQ motifs in addition to interacting via previously reported sequences. These results are discussed in terms of the function of IQGAP1 and IQ motif recognition.

Keywords

Calcium ions Cytoskeleton EF-hand IQ motif Mlc1sa Native gel 

Abbreviations

APC

Adenomatous polyposis coli

DIEA

N,N-diisopropylethylamine

FhCaM2

Fasciola hepatica type 2 calmodulin

Fmoc

9-Fluorenylmethoxycarbonyl

HATU

2-(1H-7-azabenzotriazol-1-yl)-1,1,3,3-tetramethyl uronium hexafluorophosphate methanaminium

IQ1, IQ2, IQ3, IQ4

The first, second, third and fourth IQ motifs (as numbered from the N-terminus), respectively, of human IQGAP1

Kd,app

The apparent disassociation constant under the conditions of native gel electrophoresis

LB

Luria-Bertani

TFA

Trifluoroacetic acid

VEGFR2

Vascular endothelial growth factor receptor

Notes

Acknowledgements

We thank Sean Russell for providing FhCaM2 protein which was used as a control in these experiments and Maelíosa Mc Crudden for preliminary work on the purification of Mlc1sa. This work was funded in part by a grant from the BBSRC (reference BB/D000394/1). Sevvel Pathmanathan gratefully acknowledges a studentship from Queen’s University, Belfast and the University of Jaffna (Sri Lanka) for giving him leave of absence to complete this work.

References

  1. 1.
    Hart MJ, Callow MG, Souza B, Polakis P (1996) IQGAP1, a calmodulin-binding protein with a rasGAP-related domain, is a potential effector for cdc42Hs. EMBO J 15:2997–3005PubMedGoogle Scholar
  2. 2.
    McCallum SJ, Wu WJ, Cerione RA (1996) Identification of a putative effector for Cdc42Hs with high sequence similarity to the RasGAP-related protein IQGAP1 and a Cdc42Hs binding partner with similarity to IQGAP2. J Biol Chem 271:21732–21737. doi: 10.1074/jbc.271.36.21732 PubMedCrossRefGoogle Scholar
  3. 3.
    Jeong HW, Li Z, Brown MD, Sacks DB (2007) IQGAP1 binds Rap1 and modulates its activity. J Biol Chem 282:20752–20762. doi: 10.1074/jbc.M700487200 PubMedCrossRefGoogle Scholar
  4. 4.
    Cheney RE, Mooseker MS (1992) Unconventional myosins. Curr Opin Cell Biol 4:27–35. doi: 10.1016/0955-0674(92)90055-H PubMedCrossRefGoogle Scholar
  5. 5.
    Ikura M, Clore GM, Gronenborn AM, Zhu G, Klee CB, Bax A (1992) Solution structure of a calmodulin-target peptide complex by multidimensional NMR. Science 256:632–638. doi: 10.1126/science.1585175 PubMedCrossRefGoogle Scholar
  6. 6.
    Briggs MW, Sacks DB (2003) IQGAP1 as signal integrator: Ca2+, calmodulin, Cdc42 and the cytoskeleton. FEBS Lett 542:7–11. doi: 10.1016/S0014-5793(03)00333-8 PubMedCrossRefGoogle Scholar
  7. 7.
    Machesky LM (1998) Cytokinesis: IQGAPs find a function. Curr Biol 8:R202–R205. doi: 10.1016/S0960-9822(98)70125-3 PubMedCrossRefGoogle Scholar
  8. 8.
    Weissbach L, Settleman J, Kalady MF, Snijders AJ, Murthy AE, Yan YX et al (1994) Identification of a human rasGAP-related protein containing calmodulin-binding motifs. J Biol Chem 269:20517–20521PubMedGoogle Scholar
  9. 9.
    Brill S, Li S, Lyman CW, Church DM, Wasmuth JJ, Weissbach L et al (1996) The Ras GTPase-activating-protein-related human protein IQGAP2 harbors a potential actin binding domain and interacts with calmodulin and Rho family GTPases. Mol Cell Biol 16:4869–4878PubMedGoogle Scholar
  10. 10.
    Li Q, Stuenkel EL (2004) Calcium negatively modulates calmodulin interaction with IQGAP1. Biochem Biophys Res Commun 317:787–795. doi: 10.1016/j.bbrc.2004.03.119 PubMedCrossRefGoogle Scholar
  11. 11.
    Bashour AM, Fullerton AT, Hart MJ, Bloom GS (1997) IQGAP1, a Rac- and Cdc42-binding protein, directly binds and cross-links microfilaments. J Cell Biol 137:1555–1566. doi: 10.1083/jcb.137.7.1555 PubMedCrossRefGoogle Scholar
  12. 12.
    Joyal JL, Annan RS, Ho YD, Huddleston ME, Carr SA, Hart MJ et al (1997) Calmodulin modulates the interaction between IQGAP1 and Cdc42 Identification of IQGAP1 by nanoelectrospray tandem mass spectrometry. J Biol Chem 272:15419–15425. doi: 10.1074/jbc.272.24.15419 PubMedCrossRefGoogle Scholar
  13. 13.
    Erickson JW, Cerione RA, Hart MJ (1997) Identification of an actin cytoskeletal complex that includes IQGAP and the Cdc42 GTPase. J Biol Chem 272:24443–24447. doi: 10.1074/jbc.272.39.24443 PubMedCrossRefGoogle Scholar
  14. 14.
    Li Z, Sacks DB (2003) Elucidation of the interaction of calmodulin with the IQ motifs of IQGAP1. J Biol Chem 278:4347–4352. doi: 10.1074/jbc.M208579200 PubMedCrossRefGoogle Scholar
  15. 15.
    Kuroda S, Fukata M, Kobayashi K, Nakafuku M, Nomura N, Iwamatsu A et al (1996) Identification of IQGAP as a putative target for the small GTPases, Cdc42 and Rac1. J Biol Chem 271:23363–23367. doi: 10.1074/jbc.271.38.23363 PubMedCrossRefGoogle Scholar
  16. 16.
    Mateer SC, McDaniel AE, Nicolas V, Habermacher GM, Lin MJ, Cromer DA et al (2002) The mechanism for regulation of the F-actin binding activity of IQGAP1 by calcium/calmodulin. J Biol Chem 277:12324–12333. doi: 10.1074/jbc.M109535200 PubMedCrossRefGoogle Scholar
  17. 17.
    Mateer SC, Morris LE, Cromer DA, Bensenor LB, Bloom GS (2004) Actin filament binding by a monomeric IQGAP1 fragment with a single calponin homology domain. Cell Motil Cytoskeleton 58:231–241. doi: 10.1002/cm.20013 PubMedCrossRefGoogle Scholar
  18. 18.
    Mataraza JM, Briggs MW, Li Z, Frank R, Sacks DB (2003) Identification and characterization of the Cdc42-binding site of IQGAP1. Biochem Biophys Res Commun 305:315–321. doi: 10.1016/S0006-291X(03)00759-9 PubMedCrossRefGoogle Scholar
  19. 19.
    Owen D, Campbell LJ, Littlefield K, Evetts KA, Li Z, Sacks DB et al (2008) The IQGAP1-Rac1 and IQGAP1-Cdc42 interactions: interfaces differ between the complexes. J Biol Chem 283:1692–1704. doi: 10.1074/jbc.M707257200 PubMedCrossRefGoogle Scholar
  20. 20.
    Weissbach L, Bernards A, Herion DW (1998) Binding of myosin essential light chain to the cytoskeleton-associated protein IQGAP1. Biochem Biophys Res Commun 251:269–276. doi: 10.1006/bbrc.1998.9371 PubMedCrossRefGoogle Scholar
  21. 21.
    Mbele GO, Deloulme JC, Gentil BJ, Delphin C, Ferro M, Garin J et al (2002) The zinc- and calcium-binding S100B interacts and co-localizes with IQGAP1 during dynamic rearrangement of cell membranes. J Biol Chem 277:49998–50007. doi: 10.1074/jbc.M205363200 PubMedCrossRefGoogle Scholar
  22. 22.
    Fukata M, Watanabe T, Noritake J, Nakagawa M, Yamaga M, Kuroda S et al (2002) Rac1 and Cdc42 capture microtubules through IQGAP1 and CLIP-170. Cell 109:873–885. doi: 10.1016/S0092-8674(02)00800-0 PubMedCrossRefGoogle Scholar
  23. 23.
    Ren JG, Li Z, Sacks DB (2007) IQGAP1 modulates activation of B-Raf. Proc Natl Acad Sci USA 104:10465–10469. doi: 10.1073/pnas.0611308104 PubMedCrossRefGoogle Scholar
  24. 24.
    Roy M, Li Z, Sacks DB (2004) IQGAP1 binds ERK2 and modulates its activity. J Biol Chem 279:17329–17337. doi: 10.1074/jbc.M308405200 PubMedCrossRefGoogle Scholar
  25. 25.
    Roy M, Li Z, Sacks DB (2005) IQGAP1 is a scaffold for mitogen-activated protein kinase signaling. Mol Cell Biol 25:7940–7952. doi: 10.1128/MCB.25.18.7940-7952.2005 PubMedCrossRefGoogle Scholar
  26. 26.
    Nauert JB, Rigas JD, Lester LB (2003) Identification of an IQGAP1/AKAP79 complex in beta-cells. J Cell Biochem 90:97–108. doi: 10.1002/jcb.10604 PubMedCrossRefGoogle Scholar
  27. 27.
    Yamaoka-Tojo M, Ushio-Fukai M, Hilenski L, Dikalov SI, Chen YE, Tojo T et al (2004) IQGAP1, a novel vascular endothelial growth factor receptor binding protein, is involved in reactive oxygen species-dependent endothelial migration and proliferation. Circ Res 95:276–283. doi: 10.1161/01.RES.0000136522.58649.60 PubMedCrossRefGoogle Scholar
  28. 28.
    Watanabe T, Wang S, Noritake J, Sato K, Fukata M, Takefuji M et al (2004) Interaction with IQGAP1 links APC to Rac1, Cdc42, and actin filaments during cell polarization and migration. Dev Cell 7:871–883. doi: 10.1016/j.devcel.2004.10.017 PubMedCrossRefGoogle Scholar
  29. 29.
    Le Clainche C, Schlaepfer D, Ferrari A, Klingauf M, Grohmanova K, Veligodskiy A et al (2007) IQGAP1 stimulates actin assembly through the N-WASP-Arp2/3 pathway. J Biol Chem 282:426–435. doi: 10.1074/jbc.M607711200 PubMedCrossRefGoogle Scholar
  30. 30.
    Kuroda S, Fukata M, Nakagawa M, Fujii K, Nakamura T, Ookubo T et al (1998) Role of IQGAP1, a target of the small GTPases Cdc42 and Rac1, in regulation of E-cadherin-mediated cell-cell adhesion. Science 281:832–835. doi: 10.1126/science.281.5378.832 PubMedCrossRefGoogle Scholar
  31. 31.
    Li Z, Kim SH, Higgins JM, Brenner MB, Sacks DB (1999) IQGAP1 and calmodulin modulate E-cadherin function. J Biol Chem 274:37885–37892. doi: 10.1074/jbc.274.53.37885 PubMedCrossRefGoogle Scholar
  32. 32.
    Briggs MW, Li Z, Sacks DB (2002) IQGAP1-mediated stimulation of transcriptional co-activation by beta-catenin is modulated by calmodulin. J Biol Chem 277:7453–7465. doi: 10.1074/jbc.M104315200 PubMedCrossRefGoogle Scholar
  33. 33.
    Leung J, Yueh A, Appah FS Jr, Yuan B, de los Santos K, Goff SP (2006) Interaction of Moloney murine leukemia virus matrix protein with IQGAP. EMBO J 25:2155–2166. doi: 10.1038/sj.emboj.7601097 PubMedCrossRefGoogle Scholar
  34. 34.
    Wang H, Huo R, Xu M, Lu L, Xu Z, Li J et al (2004) Cloning and characterization of a novel transcript variant of IQGAP2 in human testis. DNA Seq 15:319–325. doi: 10.1080/10425170400009012 PubMedGoogle Scholar
  35. 35.
    Wang S, Watanabe T, Noritake J, Fukata M, Yoshimura T, Itoh N et al (2007) IQGAP3, a novel effector of Rac1 and Cdc42, regulates neurite outgrowth. J Cell Sci 120:567–577. doi: 10.1242/jcs.03356 PubMedCrossRefGoogle Scholar
  36. 36.
    Lippincott J, Li R (1998) Sequential assembly of myosin II, an IQGAP-like protein, and filamentous actin to a ring structure involved in budding yeast cytokinesis. J Cell Biol 140:355–366. doi: 10.1083/jcb.140.2.355 PubMedCrossRefGoogle Scholar
  37. 37.
    Boyne JR, Yosuf HM, Bieganowski P, Brenner C, Price C (2000) Yeast myosin light chain, Mlc1p, interacts with both IQGAP and class II myosin to effect cytokinesis. J Cell Sci 113(Pt 24):4533–4543PubMedGoogle Scholar
  38. 38.
    Wendland J, Philippsen P (2002) An IQGAP-related protein, encoded by AgCYK1, is required for septation in the filamentous fungus Ashbya gossypii. Fungal Genet Biol 37:81–88. doi: 10.1016/S1087-1845(02)00034-8 PubMedCrossRefGoogle Scholar
  39. 39.
    Hailstones DL, Gunning PW (1990) Characterization of human myosin light chains 1sa and 3nm: implications for isoform evolution and function. Mol Cell Biol 10:1095–1104PubMedGoogle Scholar
  40. 40.
    Bagshaw CR (1977) On the location of the divalent metal binding sites and the light chain subunits of vertebrate myosin. Biochemistry 16:59–67. doi: 10.1021/bi00620a010 PubMedCrossRefGoogle Scholar
  41. 41.
    Hardwicke PM, Huvos P (1988) A hydrophobic region on myosin light chains modulated by divalent cations. Biochim Biophys Acta 957:352–358PubMedGoogle Scholar
  42. 42.
    Wikman-Coffelt J (1980) Properties of the non-specific calcium-binding sites of rabbit skeletal-muscle myosin. Biochem J 185:265–268PubMedGoogle Scholar
  43. 43.
    Terrak M, Wu G, Stafford WF, Lu RC, Dominguez R (2003) Two distinct myosin light chain structures are induced by specific variations within the bound IQ motifs-functional implications. EMBO J 22:362–371. doi: 10.1093/emboj/cdg058 PubMedCrossRefGoogle Scholar
  44. 44.
    Timson DJ (2003) Fine tuning the myosin motor: the role of the essential light chain in striated muscle myosin. Biochimie 85:639–645. doi: 10.1016/S0300-9084(03)00131-7 PubMedCrossRefGoogle Scholar
  45. 45.
    Timson DJ, Trayer HR, Trayer IP (1998) The N-terminus of A1-type myosin essential light chains binds actin and modulates myosin motor function. Eur J Biochem 255:654–662. doi: 10.1046/j.1432-1327.1998.2550654.x PubMedCrossRefGoogle Scholar
  46. 46.
    Timson DJ, Trayer IP (1997) The role of the proline-rich region in A1-type myosin essential light chains: implications for information transmission in the actomyosin complex. FEBS Lett 400:31–36. doi: 10.1016/S0014-5793(96)01314-2 PubMedCrossRefGoogle Scholar
  47. 47.
    Shannon KB, Li R (1999) The multiple roles of Cyk1p in the assembly and function of the actomyosin ring in budding yeast. Mol Biol Cell 10:283–296PubMedGoogle Scholar
  48. 48.
    Osman MA, Cerione RA (1998) Iqg1p, a yeast homologue of the mammalian IQGAPs, mediates cdc42p effects on the actin cytoskeleton. J Cell Biol 142:443–455. doi: 10.1083/jcb.142.2.443 PubMedCrossRefGoogle Scholar
  49. 49.
    Osman MA, Konopka JB, Cerione RA (2002) Iqg1p links spatial and secretion landmarks to polarity and cytokinesis. J Cell Biol 159:601–611. doi: 10.1083/jcb.200205084 PubMedCrossRefGoogle Scholar
  50. 50.
    Korinek WS, Bi E, Epp JA, Wang L, Ho J, Chant J (2000) Cyk3, a novel SH3-domain protein, affects cytokinesis in yeast. Curr Biol 10:947–950. doi: 10.1016/S0960-9822(00)00626-6 PubMedCrossRefGoogle Scholar
  51. 51.
    Shannon KB, Li R (2000) A myosin light chain mediates the localization of the budding yeast IQGAP-like protein during contractile ring formation. Curr Biol 10:727–730. doi: 10.1016/S0960-9822(00)00539-X PubMedCrossRefGoogle Scholar
  52. 52.
    Santamaria-Kisiel L, Rintala-Dempsey AC, Shaw GS (2006) Calcium-dependent and -independent interactions of the S100 protein family. Biochem J 396:201–214. doi: 10.1042/BJ20060195 PubMedCrossRefGoogle Scholar
  53. 53.
    Haglid KG, Hamberger A, Hansson HA, Hyden H, Persson L, Ronnback L (1975) Immunohistochemical localisation of S-100 protein in brain. Nature 258:748–749. doi: 10.1038/258748a0 PubMedCrossRefGoogle Scholar
  54. 54.
    Lennon G, Auffray C, Polymeropoulos M, Soares MB (1996) The I.M.A.G.E. consortium: an integrated molecular analysis of genomes and their expression. Genomics 33:151–152. doi: 10.1006/geno.1996.0177 PubMedCrossRefGoogle Scholar
  55. 55.
    Wu Y, Reece RJ, Ptashne M (1996) Quantitation of putative activator-target affinities predicts transcriptional activating potentials. EMBO J 15:3951–3963PubMedGoogle Scholar
  56. 56.
    Aslanidis C, de Jong PJ (1990) Ligation-independent cloning of PCR products (LIC-PCR). Nucleic Acids Res 18:6069–6074. doi: 10.1093/nar/18.20.6069 PubMedCrossRefGoogle Scholar
  57. 57.
    Bradford MM (1976) A rapid and sensitive method for the quantitation of microgram quantities of protein utilizing the principle of protein-dye binding. Anal Biochem 72:248–254. doi: 10.1016/0003-2697(76)90527-3 PubMedCrossRefGoogle Scholar
  58. 58.
    Russell SL, McFerran NV, Hoey EM, Trudgett A, Timson DJ (2007) Characterisation of two calmodulin-like proteins from the liver fluke, Fasciola hepatica. Biol Chem 388:593–599. doi: 10.1515/BC.2007.076 PubMedCrossRefGoogle Scholar
  59. 59.
    Timson DJ (2005) Functional analysis of disease-causing mutations in human UDP-galactose 4-epimerase. FEBS J 272:6170–6177. doi: 10.1111/j.1742-4658.2005.05017.x PubMedCrossRefGoogle Scholar
  60. 60.
    Fields GB, Noble RL (1990) Solid phase peptide synthesis utilizing 9-fluorenylmethoxycarbonyl amino acids. Int J Pept Protein Res 35:161–214PubMedGoogle Scholar
  61. 61.
    Marquardt D (1963) An algorithm for least squares estimation of nonlinear parameters. SIAM J Appl Math 11:431–441. doi: 10.1137/0111030 CrossRefGoogle Scholar
  62. 62.
    Rayment I, Rypniewski WR, Schmidt-Base K, Smith R, Tomchick DR, Benning MM et al (1993) Three-dimensional structure of myosin subfragment-1: a molecular motor. Science 261:50–58. doi: 10.1126/science.8316857 PubMedCrossRefGoogle Scholar
  63. 63.
    Martin SR, Bayley PM (2004) Calmodulin bridging of IQ motifs in myosin-V. FEBS Lett 567:166–170. doi: 10.1016/j.febslet.2004.04.053 PubMedCrossRefGoogle Scholar
  64. 64.
    Ren JG, Li Z, Crimmins DL, Sacks DB (2005) Self-association of IQGAP1: characterization and functional sequelae. J Biol Chem 280:34548–34557. doi: 10.1074/jbc.M507321200 PubMedCrossRefGoogle Scholar
  65. 65.
    Gillespie PG, Cyr JL (2002) Calmodulin binding to recombinant myosin-1c and myosin-1c IQ peptides. BMC Biochem 3:31. doi: 10.1186/1471-2091-3-31 PubMedCrossRefGoogle Scholar
  66. 66.
    Dickerson JB, Morgan MA, Mishra A, Slaughter CA, Morgan JI, Zheng J (2006) The influence of phosphorylation on the activity and structure of the neuronal IQ motif protein, PEP-19. Brain Res 1092:16–27. doi: 10.1016/j.brainres.2006.03.048 PubMedCrossRefGoogle Scholar
  67. 67.
    Slemmon JR, Morgan JI, Fullerton SM, Danho W, Hilbush BS, Wengenack TM (1996) Camstatins are peptide antagonists of calmodulin based upon a conserved structural motif in PEP-19, neurogranin, and neuromodulin. J Biol Chem 271:15911–15917. doi: 10.1074/jbc.271.27.15911 PubMedCrossRefGoogle Scholar
  68. 68.
    Putkey JA, Waxham MN, Gaertner TR, Brewer KJ, Goldsmith M, Kubota Y et al (2008) Acidic/IQ motif regulator of calmodulin. J Biol Chem 283:1401–1410. doi: 10.1074/jbc.M703831200 PubMedCrossRefGoogle Scholar
  69. 69.
    Theoharis NT, Sorensen BR, Theisen-Toupal J, Shea MA (2008) The neuronal voltage-dependent sodium channel type II IQ motif lowers the calcium affinity of the C-domain of calmodulin. Biochemistry 47:112–123. doi: 10.1021/bi7013129 PubMedCrossRefGoogle Scholar
  70. 70.
    Zuhlke RD, Pitt GS, Tsien RW, Reuter H (2000) Ca2+-sensitive inactivation and facilitation of L-type Ca2+ channels both depend on specific amino acid residues in a consensus calmodulin-binding motif in the α1C subunit. J Biol Chem 275:21121–21129. doi: 10.1074/jbc.M002986200 PubMedCrossRefGoogle Scholar
  71. 71.
    Coureux PD, Wells AL, Menetrey J, Yengo CM, Morris CA, Sweeney HL et al (2003) A structural state of the myosin V motor without bound nucleotide. Nature 425:419–423. doi: 10.1038/nature01927 PubMedCrossRefGoogle Scholar
  72. 72.
    Chenna R, Sugawara H, Koike T, Lopez R, Gibson TJ, Higgins DG et al (2003) Multiple sequence alignment with the Clustal series of programs. Nucleic Acids Res 31:3497–3500. doi: 10.1093/nar/gkg500 PubMedCrossRefGoogle Scholar
  73. 73.
    Bhattacharya S, Large E, Heizmann CW, Hemmings B, Chazin WJ (2003) Structure of the Ca2+/S100B/NDR kinase peptide complex: insights into S100 target specificity and activation of the kinase. Biochemistry 42:14416–14426. doi: 10.1021/bi035089a PubMedCrossRefGoogle Scholar
  74. 74.
    Inman KG, Yang R, Rustandi RR, Miller KE, Baldisseri DM, Weber DJ (2002) Solution NMR structure of S100B bound to the high-affinity target peptide TRTK-12. J Mol Biol 324:1003–1014. doi: 10.1016/S0022-2836(02)01152-X PubMedCrossRefGoogle Scholar
  75. 75.
    McClintock KA, Shaw GS (2000) A logical sequence search for S100B target proteins. Protein Sci 9:2043–2046PubMedCrossRefGoogle Scholar

Copyright information

© Springer Science+Business Media, LLC. 2008

Authors and Affiliations

  • Sevvel Pathmanathan
    • 1
  • Sarah F. Elliott
    • 1
  • Sara McSwiggen
    • 1
  • Brett Greer
    • 1
  • Pat Harriott
    • 1
  • G. Brent Irvine
    • 2
  • David J. Timson
    • 1
  1. 1.Medical Biology Centre, School of Biological SciencesQueen’s University BelfastBelfastUK
  2. 2.School of Medicine & DentistryQueen’s University BelfastBelfastUK

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