Src homology 2 domain-containing leukocyte phosphoprotein of 76 kDa and phospholipase C gamma 1 are required for NF-kappa B activation and lipid raft recruitment of protein kinase C theta induced by T cell costimulation

J Immunol. 2003 Jan 1;170(1):365-72. doi: 10.4049/jimmunol.170.1.365.

Abstract

The NF-kappaB activation pathway induced by T cell costimulation uses various molecules including Vav1 and protein kinase C (PKC)theta. Because Vav1 inducibly associates with further proteins including phospholipase C (PLC)gamma1 and Src homology 2 domain-containing leukocyte phosphoprotein of 76 kDa (SLP-76), we investigated their role for NF-kappaB activation in Jurkat leukemia T cell lines deficient for expression of these two proteins. Cells lacking SLP-76 or PLCgamma1 failed to activate NF-kappaB in response to T cell costimulation. In contrast, replenishment of SLP-76 or PLCgamma1 expression restored CD3/CD28-induced IkappaB kinase (IKK) activity as well as NF-kappaB DNA binding and transactivation. PKCtheta activated NF-kappaB in SLP-76- and PLCgamma1-deficient cells, showing that PKCtheta is acting further downstream. In contrast, Vav1-induced NF-kappaB activation was normal in SLP-76(-) cells, but absent in PLCgamma1(-) cells. CD3/CD28-stimulated recruitment of PKCtheta and IKKgamma to lipid rafts was lost in SLP-76- or PLCgamma1-negative cells, while translocation of Vav1 remained unaffected. Accordingly, recruitment of PKCtheta to the immunological synapse strictly relied on the presence of SLP-76 and PLCgamma1, but synapse translocation of Vav1 identified in this study was independent from both proteins. These results show the importance of SLP-76 and PLCgamma1 for NF-kappaB activation and raft translocation of PKCtheta and IKKgamma.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Adaptor Proteins, Signal Transducing
  • CD28 Antigens / pharmacology
  • CD3 Complex / pharmacology
  • Cell Cycle Proteins*
  • Humans
  • I-kappa B Kinase
  • Isoenzymes / biosynthesis
  • Isoenzymes / genetics
  • Isoenzymes / metabolism*
  • Isoenzymes / physiology*
  • Jurkat Cells
  • Lymphocyte Activation* / genetics
  • Membrane Microdomains / enzymology*
  • Membrane Microdomains / genetics
  • NF-kappa B / biosynthesis
  • NF-kappa B / metabolism*
  • Phospholipase C gamma
  • Phosphoproteins / deficiency
  • Phosphoproteins / genetics
  • Phosphoproteins / physiology*
  • Protein Kinase C / biosynthesis
  • Protein Kinase C / genetics
  • Protein Kinase C / metabolism*
  • Protein Kinase C-theta
  • Protein Serine-Threonine Kinases / metabolism
  • Protein Transport / genetics
  • Protein Transport / immunology
  • Proto-Oncogene Proteins / biosynthesis
  • Proto-Oncogene Proteins / metabolism
  • Proto-Oncogene Proteins c-vav
  • Signal Transduction / genetics
  • Signal Transduction / immunology
  • T-Lymphocytes / immunology*
  • Transfection
  • Type C Phospholipases / genetics
  • Type C Phospholipases / physiology*
  • src Homology Domains / genetics
  • src Homology Domains / immunology*

Substances

  • Adaptor Proteins, Signal Transducing
  • CD28 Antigens
  • CD3 Complex
  • Cell Cycle Proteins
  • Isoenzymes
  • NF-kappa B
  • Phosphoproteins
  • Proto-Oncogene Proteins
  • Proto-Oncogene Proteins c-vav
  • SLP-76 signal Transducing adaptor proteins
  • VAV1 protein, human
  • Protein Serine-Threonine Kinases
  • CHUK protein, human
  • I-kappa B Kinase
  • IKBKB protein, human
  • IKBKE protein, human
  • PRKCQ protein, human
  • Protein Kinase C
  • Protein Kinase C-theta
  • Type C Phospholipases
  • Phospholipase C gamma