The receptor for antigen and MHC (T3-Ti) on the vast majority of human T lymphocytes is a molecular complex comprised of an immunoglobulin-like Ti alpha-beta disulfide-linked heterodimer and three non-covalently associated monomorphic subunits (T3 gamma, T3 delta and T3 epsilon). Although much is known about the molecular nature of Ti diversity encoded by V, D and J-like elements and the primary structures of individual T3 subunits, the signal transduction process is ill-defined. Its molecular basis will be the subject of the present proposal. Firstly, monoclonal antibodies will be produced to the native external domain of individual T3 subunits expressed in the baculoviral system. These reagents will allow us to determine whether there are unique functional effects attributable to a given subunit. Alterations in (Ca2+)i, phosphoinositide turnover, phosphorylation events and gene transcription will be examined after external domain perturbation. Furthermore, novel strategies for developing antibodies to other putative T3-Ti associated surface structures will be devised and their primary structure analyzed by microsequencing and cDNA cloning. Secondly, through the use of gene transfer techniques and truncation mutants, the signal transduction function of individual cytoplasmic domains of T3 gamma, T3 delta and T3 epsilon will be defined. The role of potential phosphorylation sites in the cytoplasmic domain will be examined via site directed mutagenesis. The significance of the single acidic amino acid residue in the transmembrane segment of each T3 subunit with respect to T3-Ti stability and function will be investigated. Expression of T3-Ti subunit RNAs in xenopus oocytes will be attempted with the view of coupling T3-Ti complex signalling to the endogenous oocyte IP3 triggered C1- channel as a rapid assay for receptor mediated activation events. Thirdly, the role of the T3-Ti subunits in signalling growth inhibition will be characterized. It will be determined whether anti-T3 modulation functionally uncouples the T3-Ti complex from a G protein, inhibits protein kinase C or induces an arrestin- like protein to bind the T3-Ti complex. This desensitization response will be studied in detail with Jurkat mutants which have cytoplasmic deletions of individual T3 subunits. Finally, as the binding of the LFA-3 molecule to the CD2 external domain appears to amplify triggering through the T3-Ti complex, we will determine whether the T3 subunits are modified by phosphorylation or new protein associations after such an interaction.

Agency
National Institute of Health (NIH)
Institute
National Institute of Allergy and Infectious Diseases (NIAID)
Type
Research Project (R01)
Project #
5R01AI019807-10
Application #
3129240
Study Section
Immunobiology Study Section (IMB)
Project Start
1983-05-01
Project End
1993-11-30
Budget Start
1992-05-01
Budget End
1993-11-30
Support Year
10
Fiscal Year
1992
Total Cost
Indirect Cost
Name
Dana-Farber Cancer Institute
Department
Type
DUNS #
149617367
City
Boston
State
MA
Country
United States
Zip Code
02215
Mallis, Robert J; Arthanari, Haribabu; Lang, Matthew J et al. (2018) NMR-directed design of pre-TCR? and pMHC molecules implies a distinct geometry for pre-TCR relative to ??TCR recognition of pMHC. J Biol Chem 293:754-766
Mallis, Robert J; Reinherz, Ellis L; Wagner, Gerhard et al. (2016) Backbone resonance assignment of N15, N30 and D10 T cell receptor ? subunits. Biomol NMR Assign 10:35-9
Reinherz, Ellis L; Wang, Jia-huai (2015) Codification of bidentate pMHC interaction with TCR and its co-receptor. Trends Immunol 36:300-6
Mallis, Robert J; Bai, Ke; Arthanari, Haribabu et al. (2015) Pre-TCR ligand binding impacts thymocyte development before ??TCR expression. Proc Natl Acad Sci U S A 112:8373-8
Choi, Young I; Duke-Cohan, Jonathan S; Chen, Wei et al. (2014) Dynamic control of ?1 integrin adhesion by the plexinD1-sema3E axis. Proc Natl Acad Sci U S A 111:379-84
Choi, Young I; Duke-Cohan, Jonathan S; Tan, Jing et al. (2013) Plxnd1 expression in thymocytes regulates their intrathymic migration while that in thymic endothelium impacts medullary topology. Front Immunol 4:392
Touma, Maki; Keskin, Derin B; Shiroki, Fumiko et al. (2011) Impaired B cell development and function in the absence of IkappaBNS. J Immunol 187:3942-52
Kim, Sun Taek; Touma, Maki; Takeuchi, Koh et al. (2010) Distinctive CD3 heterodimeric ectodomain topologies maximize antigen-triggered activation of alpha beta T cell receptors. J Immunol 185:2951-9
Vernier, Gregory; Wang, Jie; Jennings, Laura D et al. (2009) Solubilization and characterization of the anthrax toxin pore in detergent micelles. Protein Sci 18:1882-95
Kim, Sun Taek; Takeuchi, Koh; Sun, Zhen-Yu J et al. (2009) The alphabeta T cell receptor is an anisotropic mechanosensor. J Biol Chem 284:31028-37

Showing the most recent 10 out of 104 publications