Dynamin 2 regulates T cell activation by controlling actin polymerization at the immunological synapse


Actin reorganization at the immunological synapse is required for the amplification and generation of a functional immune response. Using small interfering RNA, we show here that dynamin 2 (Dyn2), a large GTPase involved in receptor-mediated internalization, did not alter antibody-mediated T cell receptor internalization but considerably affected T cell receptor–stimulated T cell activation by regulating multiple biochemical signaling pathways and the accumulation of F-actin at the immunological synapse. Moreover, Dyn2 interacted directly with the Rho family guanine nucleotide exchange factor Vav1, and this interaction was required for T cell activation. These data identify a functionally important interaction between Dyn2 and Vav1 that regulates actin reorganization and multiple signaling pathways in T lymphocytes.

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Figure 1: Dyn2 is recruited to the T cell–APC contact site.
Figure 2: Dyn2 regulates T cell activation.
Figure 3: TCR expression and antibody-mediated internalization are not altered in Dyn2 suppressed cells.
Figure 4: Dynamin2 suppression inhibits actin polarization at the T cell–APC contact site.
Figure 5: The Dyn2 PRD is required for T cell activation.
Figure 6: The Dyn2 PRD specifically, directly and functionally interacts with the C-terminal SH3 domain of Vav1.
Figure 7: Dyn2 couples TCR signaling to the activation of Erk, Jnk and PLC-γ1.


  1. 1

    Holsinger, L.J. et al. Defects in actin-cap formation in Vav-deficient mice implicate an actin requirement for lymphocyte signal transduction. Curr. Biol. 8, 563–572 (1998).

    CAS  Article  Google Scholar 

  2. 2

    Morgan, M.M. et al. Superantigen-induced T cell:B cell conjugation is mediated by LFA-1 and requires signaling through Lck, but not ZAP-70. J. Immunol. 167, 5708–5718 (2001).

    CAS  Article  Google Scholar 

  3. 3

    Bubeck Wardenburg, J. et al. Regulation of PAK activation and the T cell cytoskeleton by the linker protein SLP-76. Immunity 9, 607–616 (1998).

    CAS  Article  Google Scholar 

  4. 4

    Bunnell, S.C., Kapoor, V., Trible, R.P., Zhang, W. & Samelson, L.E. Dynamic actin polymerization drives T cell receptor-induced spreading: a role for the signal transduction adaptor LAT. Immunity 14, 315–329 (2001).

    CAS  Article  Google Scholar 

  5. 5

    Cannon, J.L. & Burkhardt, J.K. The regulation of actin remodeling during T-cell-APC conjugate formation. Immunol. Rev. 186, 90–99 (2002).

    CAS  Article  Google Scholar 

  6. 6

    Stowers, L., Yelon, D., Berg, L.J. & Chant, J. Regulation of the polarization of T cells toward antigen-presenting cells by Ras-related GTPase CDC42. Proc. Natl. Acad. Sci. USA 92, 5027–5031 (1995).

    CAS  Article  Google Scholar 

  7. 7

    Zeng, R. et al. SLP-76 coordinates Nck-dependent Wiskott-Aldrich syndrome protein recruitment with Vav-1/Cdc42-dependent Wiskott-Aldrich syndrome protein activation at the T cell-APC contact site. J. Immunol. 171, 1360–1368 (2003).

    CAS  Article  Google Scholar 

  8. 8

    Krawczyk, C. et al. Vav1 controls integrin clustering and MHC/peptide-specific cell adhesion to antigen-presenting cells. Immunity 16, 331–343 (2002).

    CAS  Article  Google Scholar 

  9. 9

    Snapper, S.B. et al. Wiskott-Aldrich syndrome protein-deficient mice reveal a role for WASP in T but not B cell activation. Immunity 9, 81–91 (1998).

    CAS  Article  Google Scholar 

  10. 10

    Labno, C.M. et al. Itk functions to control actin polymerization at the immune synapse through localized activation of Cdc42 and WASP. Curr. Biol. 13, 1619–1624 (2003).

    CAS  Article  Google Scholar 

  11. 11

    Orth, J.D. & McNiven, M.A. Dynamin at the actin-membrane interface. Curr. Opin. Cell Biol. 15, 31–39 (2003).

    CAS  Article  Google Scholar 

  12. 12

    Praefcke, G.J. & McMahon, H.T. The dynamin superfamily: universal membrane tubulation and fission molecules? Nat. Rev. Mol. Cell Biol. 5, 133–147 (2004).

    CAS  Article  Google Scholar 

  13. 13

    Cao, Y. et al. Pleiotropic defects in TCR signaling in a Vav-1-null Jurkat T-cell line. EMBO J. 21, 4809–4819 (2002).

    CAS  Article  Google Scholar 

  14. 14

    Zakaria, S. et al. Differential regulation of TCR-mediated gene transcription by Vav family members. J. Exp. Med. 199, 429–434 (2004).

    CAS  Article  Google Scholar 

  15. 15

    Gout, I. et al. The GTPase dynamin binds to and is activated by a subset of SH3 domains. Cell 75, 25–36 (1993).

    CAS  Article  Google Scholar 

  16. 16

    Damke, H., Baba, T., Warnock, D.E. & Schmid, S.L. Induction of mutant dynamin specifically blocks endocytic coated vesicle formation. J. Cell Biol. 127, 915–934 (1994).

    CAS  Article  Google Scholar 

  17. 17

    Okamoto, P.M., Herskovits, J.S. & Vallee, R.B. Role of the basic, proline-rich region of dynamin in Src homology 3 domain binding and endocytosis. J. Biol. Chem. 272, 11629–11635 (1997).

    CAS  Article  Google Scholar 

  18. 18

    So, C.W. et al. The interaction between EEN and Abi-1, two MLL fusion partners, and synaptojanin and dynamin: implications for leukaemogenesis. Leukemia 14, 594–601 (2000).

    CAS  Article  Google Scholar 

  19. 19

    Turner, M. & Billadeau, D.D. VAV proteins as signal integrators for multi-subunit immune-recognition receptors. Nat. Rev. Immunol. 2, 476–486 (2002).

    CAS  Article  Google Scholar 

  20. 20

    Shajahan, A.N. et al. Role of Src-induced dynamin-2 phosphorylation in caveolae-mediated endocytosis in endothelial cells. J. Biol. Chem. 279, 20392–20400 (2004).

    CAS  Article  Google Scholar 

  21. 21

    McNiven, M.A. et al. Regulated interactions between dynamin and the actin-binding protein cortactin modulate cell shape. J. Cell Biol. 151, 187–198 (2000).

    CAS  Article  Google Scholar 

  22. 22

    Houlard, M. et al. Vav1 is a component of transcriptionally active complexes. J. Exp. Med. 195, 1115–1127 (2002).

    CAS  Article  Google Scholar 

  23. 23

    Zugaza, J.L. et al. Structural determinants for the biological activity of Vav proteins. J. Biol. Chem. 277, 45377–45392 (2002).

    CAS  Article  Google Scholar 

  24. 24

    Cao, Y. et al. Pleiotropic defects in TCR signaling in a Vav-1-null Jurkat T-cell line. EMBO J. 21, 4809–4819 (2002).

    CAS  Article  Google Scholar 

  25. 25

    Costello, P.S. et al. The Rho-family GTP exchange factor Vav is a critical transducer of T cell receptor signals to the calcium, ERK, and NF-κB pathways. Proc. Natl. Acad. Sci. USA 96, 3035–3040 (1999).

    CAS  Article  Google Scholar 

  26. 26

    Reynolds, L.F. et al. Vav1 transduces T cell receptor signals to the activation of phospholipase C-γ1 via phosphoinositide 3-kinase-dependent and -independent pathways. J. Exp. Med. 195, 1103–1114 (2002).

    CAS  Article  Google Scholar 

  27. 27

    Reynolds, L.F. et al. Vav1 transduces T cell receptor signals to the activation of the Ras/ERK pathway via LAT, Sos, and RasGRP1. J. Biol. Chem. 279, 18239–18246 (2004).

    CAS  Article  Google Scholar 

  28. 28

    Schlunck, G. et al. Modulation of Rac localization and function by dynamin. Mol. Biol. Cell 15, 256–267 (2004).

    CAS  Article  Google Scholar 

  29. 29

    Seedorf, K. et al. Dynamin binds to SH3 domains of phospholipase C gamma and GRB-2. J. Biol. Chem. 269, 16009–16014 (1994).

    CAS  PubMed  Google Scholar 

  30. 30

    Schaeffer, E.M. et al. Requirement for Tec kinases Rlk and Itk in T cell receptor signaling and immunity. Science 284, 638–641 (1999).

    CAS  Article  Google Scholar 

  31. 31

    Finkelstein, L.D. & Schwartzberg, P.L. Tec kinases: shaping T-cell activation through actin. Trends Cell Biol. 14, 443–451 (2004).

    CAS  Article  Google Scholar 

  32. 32

    Schaefer, A.W. et al. Activation of the MAPK signal cascade by the neural cell adhesion molecule L1 requires L1 internalization. J. Biol. Chem. 274, 37965–37973 (1999).

    CAS  Article  Google Scholar 

  33. 33

    Daaka, Y. et al. Essential role for G protein-coupled receptor endocytosis in the activation of mitogen-activated protein kinase. J. Biol. Chem. 273, 685–688 (1998).

    CAS  Article  Google Scholar 

  34. 34

    Kranenburg, O., Verlaan, I. & Moolenaar, W.H. Dynamin is required for the activation of mitogen-activated protein (MAP) kinase by MAP kinase kinase. J. Biol. Chem. 274, 35301–35304 (1999).

    CAS  Article  Google Scholar 

  35. 35

    Panigada, M. et al. Constitutive endocytosis and degradation of the pre-T cell receptor. J. Exp. Med. 195, 1585–1597 (2002).

    CAS  Article  Google Scholar 

  36. 36

    Rivero-Lezcano, O.M., Marcilla, A., Sameshima, J.H. & Robbins, K.C. Wiskott-Aldrich syndrome protein physically associates with Nck through Src homology 3 domains. Mol. Cell. Biol. 15, 5725–5731 (1995).

    CAS  Article  Google Scholar 

  37. 37

    Anton, I.M., Lu, W., Mayer, B.J., Ramesh, N. & Geha, R.S. The Wiskott-Aldrich syndrome protein-interacting protein (WIP) binds to the adaptor protein Nck. J. Biol. Chem. 273, 20992–20995 (1998).

    CAS  Article  Google Scholar 

  38. 38

    Krueger, E.W., Orth, J.D., Cao, H. & McNiven, M.A. A dynamin-cortactin-Arp2/3 complex mediates actin reorganization in growth factor-stimulated cells. Mol. Biol. Cell 14, 1085–1096 (2003).

    CAS  Article  Google Scholar 

  39. 39

    Taniuchi, I. et al. Antigen-receptor induced clonal expansion and deletion of lymphocytes are impaired in mice lacking HS1 protein, a substrate of the antigen-receptor-coupled tyrosine kinases. EMBO J. 14, 3664–3678 (1995).

    CAS  Article  Google Scholar 

  40. 40

    Innocenti, M. et al. Abi1 is essential for the formation and activation of a WAVE2 signalling complex. Nat. Cell Biol. 6, 319–327 (2004).

    CAS  Article  Google Scholar 

  41. 41

    Williams, B.L. et al. Genetic evidence for differential coupling of Syk family kinases to the T-cell receptor: reconstitution studies in a ZAP-70-deficient Jurkat T-cell line. Mol. Cell. Biol. 18, 1388–1399 (1998).

    CAS  Article  Google Scholar 

  42. 42

    Henley, J.R. & McNiven, M.A. Association of a dynamin-like protein with the Golgi apparatus in mammalian cells. J. Cell Biol. 133, 761–775 (1996).

    CAS  Article  Google Scholar 

  43. 43

    Henley, J.R., Krueger, E.W., Oswald, B.J. & McNiven, M.A. Dynamin-mediated internalization of caveolae. J. Cell Biol. 141, 85–99 (1998).

    CAS  Article  Google Scholar 

  44. 44

    Ting, A.T., Karnitz, L.M., Schoon, R.A., Abraham, R.T. & Leibson, P.J. Fc gamma receptor activation induces the tyrosine phosphorylation of both phospholipase C (PLC)-γ1 and PLC-γ2 in natural killer cells. J. Exp. Med. 176, 1751–1755 (1992).

    CAS  Article  Google Scholar 

  45. 45

    Billadeau, D.D., Mackie, S.M., Schoon, R.A. & Leibson, P.J. The Rho family guanine nucleotide exchange factor Vav-2 regulates the development of cell-mediated cytotoxicity. J. Exp. Med. 192, 381–392 (2000).

    CAS  Article  Google Scholar 

  46. 46

    Jevremovic, D. et al. Cutting edge: a role for the adaptor protein LAT in human NK cell-mediated cytotoxicity. J. Immunol. 162, 2453–2456 (1999).

    CAS  Google Scholar 

  47. 47

    Trushin, S.A. et al. Protein kinase Cα (PKCα) acts upstream of PKCθ to activate IκB kinase and NF-κB in T lymphocytes. Mol. Cell. Biol. 23, 7068–7081 (2003).

    CAS  Article  Google Scholar 

  48. 48

    McKean, D.J. et al. Maturation versus death of developing double-positive thymocytes reflects competing effects on Bcl-2 expression and can be regulated by the intensity of CD28 costimulation. J. Immunol. 166, 3468–3475 (2001).

    CAS  Article  Google Scholar 

  49. 49

    Bell, M.P., Huntoon, C.J., Graham, D. & McKean, D.J. The analysis of costimulatory receptor signaling cascades in normal T lymphocytes using in vitro gene transfer and reporter gene analysis. Nat. Med. 7, 1155–1158 (2001).

    CAS  Article  Google Scholar 

  50. 50

    Cannon, J.L. et al. Wasp recruitment to the T cell:APC contact site occurs independently of Cdc42 activation. Immunity 15, 249–259 (2001).

    CAS  Article  Google Scholar 

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The authors thank P.J. Leibson for critical reading of the manuscript. Supported by the Mayo Foundation, US National Institutes of Health (AI44959 to D.J.M., and CA47752 to D.D.B.) and Cancer Research Institute (D.D.B.).

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Correspondence to Daniel D Billadeau.

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The authors declare no competing financial interests.

Supplementary information

Supplementary Fig. 1

Dyn2 suppression does not affect proximal TCR signaling events. (PDF 142 kb)

Supplementary Fig. 2

Verification of the Vav1-Dyn2 interaction. (PDF 101 kb)

Supplementary Fig. 3

PLCγ1, SLP-76 and Sos1 are effectively recruited to LAT in the absence of Dyn2. (PDF 200 kb)

Supplementary Fig. 4

Dyn2 requires Vav1 for efficient localization to the IS. (PDF 236 kb)

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Gomez, T., Hamann, M., McCarney, S. et al. Dynamin 2 regulates T cell activation by controlling actin polymerization at the immunological synapse. Nat Immunol 6, 261–270 (2005). https://doi.org/10.1038/ni1168

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