The process by which T cells are activated, and the consequences of activation (e.g. new gene transcription, cytokine production, apoptosis), are being investigated by a variety of approaches: 1) Tumor Necrosis Factor (TNF) causes many of its biological responses by activating the transcription factor NF-kB. We have found that this process involves the polyubiquitination of an intermediary adaptor protein, RIP, and subsequent recognition of this modified protein by NEMO, a critical regulator of NF-kB activity. NEMO is the first protein to be identified as a highly-specific binding partner for polyubiquitin that has internal Lys 63-linkages. 2) p38 is a MAP kinase (MAPK) involved in inflammatory processes. Like all MAPK, p38 is activated by Thr/Tyr dual phosphorylation via upstream MAP kinase kinases (MAPKK). We have found that T cells possess an alternative p38 activation pathway involving phosphorylation of a novel tyrosine residue in a MAPKK-independent manner. We have shown that mice lacking Gadd45a, an inhibitor of the alternative activation pathway, paradoxically have defective type 1 (inflammatory) T helper cell responses because p38 is not activated normally in dendritic cells. Therefore, Gadd45a has two tissue-specific opposing functions with regard to p38. 3) TNF signaling causes the recruitment of a protein, c-IAP1, that we have identified as a ubiquitin protein ligase (E3), to the signaling complex. We have shown in normal B cells that c-IAP1 ubiquitinates a critical enzyme in the MAPK cascade, ASK1, resulting in its degradation and subsequent termination of downstream MAPK signaling. Thus, TNF initiates a feedback loop that limits its own signaling pathways. 4) After an immune response, a small number of responding T cells (memory T cells) survive and persist for years, poised to respond rapidly if the original antigens are re-encountered. Although thought to be unidirectional, we have found in preliminary studies have this process is reversible, and memory T cells can be induced to assume nave phenotype and function. Work on the mechanisms by which memory is maintained is ongoing.

Agency
National Institute of Health (NIH)
Institute
Division of Basic Sciences - NCI (NCI)
Type
Intramural Research (Z01)
Project #
1Z01BC009290-21
Application #
7338163
Study Section
(LICB)
Project Start
Project End
Budget Start
Budget End
Support Year
21
Fiscal Year
2006
Total Cost
Indirect Cost
Name
Basic Sciences
Department
Type
DUNS #
City
State
Country
United States
Zip Code
Ashwell, Jonathan D (2006) The many paths to p38 mitogen-activated protein kinase activation in the immune system. Nat Rev Immunol 6:532-40
Schito, Marco L; Demidov, Oleg N; Saito, Shin'ichi et al. (2006) Wip1 phosphatase-deficient mice exhibit defective T cell maturation due to sustained p53 activation. J Immunol 176:4818-25
Wu, Chuan-Jin; Conze, Dietrich B; Li, Tao et al. (2006) Sensing of Lys 63-linked polyubiquitination by NEMO is a key event in NF-kappaB activation [corrected] Nat Cell Biol 8:398-406
Mittelstadt, Paul R; Salvador, Jesus M; Fornace Jr, Albert J et al. (2005) Activating p38 MAPK: new tricks for an old kinase. Cell Cycle 4:1189-92
Wu, Chuan-Jin; Conze, Dietrich B; Li, Xiaoming et al. (2005) TNF-alpha induced c-IAP1/TRAF2 complex translocation to a Ubc6-containing compartment and TRAF2 ubiquitination. EMBO J 24:1886-98
Munitic, Ivana; Ryan, Philip E; Ashwell, Jonathan D (2005) T cells in G1 provide a memory-like response to secondary stimulation. J Immunol 174:4010-8
Salvador, Jesus M; Mittelstadt, Paul R; Belova, Galina I et al. (2005) The autoimmune suppressor Gadd45alpha inhibits the T cell alternative p38 activation pathway. Nat Immunol 6:396-402
Salvador, Jesus M; Mittelstadt, Paul R; Guszczynski, Tad et al. (2005) Alternative p38 activation pathway mediated by T cell receptor-proximal tyrosine kinases. Nat Immunol 6:390-5
Mittelstadt, Paul R; Ashwell, Jonathan D (2003) Disruption of glucocorticoid receptor exon 2 yields a ligand-responsive C-terminal fragment that regulates gene expression. Mol Endocrinol 17:1534-42
Rengarajan, J; Mittelstadt, P R; Mages, H W et al. (2000) Sequential involvement of NFAT and Egr transcription factors in FasL regulation. Immunity 12:293-300

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