Molecules and Cells

Korean Society for Molecular and Cellular Biology (한국분자세포생물학회)
권호 표지
DOI QR코드

DOI QR Code

PKCθ-Mediated PDK1 Phosphorylation Enhances T Cell Activation by Increasing PDK1 Stability

  • Kang, Jung-Ah (School of Life Sciences and Cell Logistics Research Center, Gwangju Institute of Science and Technology (GIST)) ;
  • Choi, Hyunwoo (School of Life Sciences and Cell Logistics Research Center, Gwangju Institute of Science and Technology (GIST)) ;
  • Yang, Taewoo (School of Life Sciences and Cell Logistics Research Center, Gwangju Institute of Science and Technology (GIST)) ;
  • Cho, Steve K. (School of Life Sciences and Cell Logistics Research Center, Gwangju Institute of Science and Technology (GIST)) ;
  • Park, Zee-Yong (School of Life Sciences and Cell Logistics Research Center, Gwangju Institute of Science and Technology (GIST)) ;
  • Park, Sung-Gyoo (School of Life Sciences and Cell Logistics Research Center, Gwangju Institute of Science and Technology (GIST))
  • Received : 2016.10.05
  • Accepted : 2016.12.12
  • Published : 2017.01.31
Download PDF
⟨ Previous Next ⟩

Abstract

PDK1 is essential for T cell receptor (TCR)-mediated activation of $NF-{\kappa}B$, and PDK1-induced phosphorylation of $PKC{\theta}$ is important for TCR-induced $NF-{\kappa}B$ activation. However, inverse regulation of PDK1 by $PKC{\theta}$ during T cell activation has not been investigated. In this study, we found that $PKC{\theta}$ is involved in human PDK1 phosphorylation and that its kinase activity is crucial for human PDK1 phosphorylation. Mass spectrometry analysis of wild-type $PKC{\theta}$ or of kinase-inactive form of $PKC{\theta}$ revealed that $PKC{\theta}$ induced phosphorylation of human PDK1 at Ser-64. This $PKC{\theta}$-induced PDK1 phosphorylation positively regulated T cell activation and TCR-induced $NF-{\kappa}B$ activation. Moreover, phosphorylation of human PDK1 at Ser-64 increased the stability of human PDK1 protein. These results suggest that Ser-64 is an important phosphorylation site that is part of a positive feedback loop for human PDK1-$PKC{\theta}$-mediated T cell activation.

Keywords

References

  1. Balendran, A., Biondi, R.M., Cheung, P.C., Casamayor, A., Deak, M., and Alessi, D.R. (2000). A 3-phosphoinositide-dependent protein kinase-1 (PDK1) docking site is required for the phosphorylation of protein kinase Czeta (PKCzeta ) and PKC-related kinase 2 by PDK1. J. Biol. Chem. 275, 20806-20813. https://doi.org/10.1074/jbc.M000421200
  2. Bhatt, D., and Ghosh, S. (2014). Regulation of the NF-kappaB-mediated transcription of inflammatory genes. Front. Immunol. 5, 71.
  3. Casamayor, A., Morrice, N.A., and Alessi, D.R. (1999). Phosphorylation of Ser-241 is essential for the activity of 3-phosphoinositide-dependent protein kinase-1: identification of five sites of phosphorylation in vivo. Biochem. J. 342 (Pt 2), 287-292. https://doi.org/10.1042/bj3420287
  4. Cho, J.E., Kim, Y.S., Park, S., Cho, S.N., and Lee, H. (2010). Mycobacterium tuberculosis-induced expression of Leukotactin-1 is mediated by the PI3-K/PDK1/Akt signaling pathway. Mol. Cells 29, 35-39. https://doi.org/10.1007/s10059-010-0003-5
  5. Eckerdt, F., Yuan, J., Saxena, K., Martin, B., Kappel, S., Lindenau, C., Kramer, A., Naumann, S., Daum, S., Fischer, G., et al. (2005). Polo-like kinase 1-mediated phosphorylation stabilizes Pin1 by inhibiting its ubiquitination in human cells. J. Biol. Chem. 280, 36575-36583. https://doi.org/10.1074/jbc.M504548200
  6. Kang, J.A., Jeong, S.P., Park, D., Hayden, M.S., Ghosh, S., and Park, S.G. (2013). Transition from heterotypic to homotypic PDK1 homodimerization is essential for TCR-mediated NF-kappaB activation. J. Immunol. 190, 4508-4515. https://doi.org/10.4049/jimmunol.1202923
  7. Kenchappa, P., Yadav, A., Singh, G., Nandana, S., and Banerjee, K. (2004). Rescue of TNFalpha-inhibited neuronal cells by IGF-1 involves Akt and c-Jun N-terminal kinases. J. Neurosci. Res. 76, 466-474. https://doi.org/10.1002/jnr.20081
  8. Lee, K.Y., D'Acquisto, F., Hayden, M.S., Shim, J.H., and Ghosh, S. (2005). PDK1 nucleates T cell receptor-induced signaling complex for NF-kappaB activation. Science 308, 114-118. https://doi.org/10.1126/science.1107107
  9. Mestas, J., and Hughes, C.C. (2004). Of mice and not men: differences between mouse and human immunology. J. Immunol. 172, 2731-2738. https://doi.org/10.4049/jimmunol.172.5.2731
  10. Mora, A., Komander, D., van Aalten, D.M., and Alessi, D.R. (2004). PDK1, the master regulator of AGC kinase signal transduction. Semin. Cell Dev. Biol. 15, 161-170. https://doi.org/10.1016/j.semcdb.2003.12.022
  11. Park, S.G., Schulze-Luehrman, J., Hayden, M.S., Hashimoto, N., Ogawa, W., Kasuga, M., and Ghosh, S. (2009). The kinase PDK1 integrates T cell antigen receptor and CD28 coreceptor signaling to induce NF-kappaB and activate T cells. Nat. Immunol. 10, 158-166. https://doi.org/10.1038/ni.1687
  12. Park, S.G., Long, M., Kang, J.A., Kim, W.S., Lee, C.R., Im, S.H., Strickland, I., Schulze-Luehrmann, J., Hayden, M.S., and Ghosh, S. (2013). The kinase PDK1 is essential for B-cell receptor mediated survival signaling. PloS One 8, e55378. https://doi.org/10.1371/journal.pone.0055378
  13. Riojas, R.A., Kikani, C.K., Wang, C., Mao, X., Zhou, L., Langlais, P.R., Hu, D., Roberts, J.L., Dong, L.Q., and Liu, F. (2006). Fine tuning PDK1 activity by phosphorylation at Ser163. J. Biol. Chem. 281, 21588-21593. https://doi.org/10.1074/jbc.M600393200
  14. Scheid, M.P., Parsons, M., and Woodgett, J.R. (2005). Phosphoinositide-dependent phosphorylation of PDK1 regulates nuclear translocation. Mol. Cell. Biol. 25, 2347-2363. https://doi.org/10.1128/MCB.25.6.2347-2363.2005
  15. Seong, H.A., Jung, H., Ichijo, H., and Ha, H. (2010). Reciprocal negative regulation of PDK1 and ASK1 signaling by direct interaction and phosphorylation. J. Biol. Chem. 285, 2397-2414. https://doi.org/10.1074/jbc.M109.064295
  16. Villalba, M., Coudronniere, N., Deckert, M., Teixeiro, E., Mas, P., and Altman, A. (2000). A novel functional interaction between Vav and PKCtheta is required for TCR-induced T cell activation. Immunity 12, 151-160. https://doi.org/10.1016/S1074-7613(00)80168-5
  17. Villalba, M., Bi, K., Hu, J., Altman, Y., Bushway, P., Reits, E., Neefjes, J., Baier, G., Abraham, R.T., and Altman, A. (2002). Translocation of PKC[theta] in T cells is mediated by a nonconventional, PI3-K- and Vav-dependent pathway, but does not absolutely require phospholipase C. J. Cell. Biol. 157, 253-263. https://doi.org/10.1083/jcb.200201097
  18. Wang, C., Liu, M., Riojas, R.A., Xin, X., Gao, Z., Zeng, R., Wu, J., Dong, L.Q., and Liu, F. (2009). Protein kinase C theta (PKCtheta)-dependent phosphorylation of PDK1 at Ser504 and Ser532 contributes to palmitate-induced insulin resistance. J. Biol. Chem. 284, 2038-2044. https://doi.org/10.1074/jbc.M806336200
  19. Wang, X., Chuang, H.C., Li, J.P., and Tan, T.H. (2012). Regulation of PKC-theta function by phosphorylation in T cell receptor signaling. Front. Immunol. 3, 197.

Cited by

  1. microRNA-145-3p inhibits non-small cell lung cancer cell migration and invasion by targeting PDK1 via the mTOR signaling pathway vol.119, pp.1, 2018, https://doi.org/10.1002/jcb.26252
  2. Myeloid deletion of phosphoinositide-dependent kinase-1 enhances NK cell-mediated antitumor immunity by mediating macrophage polarization vol.9, pp.1, 2020, https://doi.org/10.1080/2162402x.2020.1774281
  3. FasL-PDPK1 Pathway Promotes the Cytotoxicity of CD8+ T Cells During Ischemic Stroke vol.11, pp.4, 2020, https://doi.org/10.1007/s12975-019-00749-0