Although most of the molecules involved in TCR signaling have likely been identified, our understanding of the basal signaling versus the induced signaling states are rather rudimentary. Tyrosine kinases and phosphatases maintain the dynamic equilibrium that controls and maintains the both the basal state and changes in their functional activities or spatial localization are important for Induced tyrosine phosphorylation. This project focuses on understanding the specificity and complex regulation of these tyrosine kinases and phosphatases on at the plasma membrane. Understanding the regulation of phosphorylation of the TCR cytoplasmic Z-chain as well as of LAT proteins will be the endpoints we will focus on. We will use: purified recombinant proteins;novel genetically controlled kinase inhibitors;a model two dimensional lipid bilayer system containing defined quantities of recombinant proteins;and, computational modeling to describe and compare the simple bilayer system to existing well-studied intact cekks or membranes of model cellular systems. We will compare the biochemical behavior of such two dimensional models to cellular systems in an iterative way, increasing component complexity and by applying computational modeling. We expect such analyses to reveal expected as well as unanticipated behaviors. We also expect the comparison of the simple system to more complex systems to reveal the presence of regulatory pathways that may not have previously been appreciated, such as positive and negative feedback circuitry. We clearly recognize that we cannot model all of the complexity of TCR signaling but we aim to understand the detailed mechanisms controlling tyrosine phosphorylation of the TCR Z-chain and how these events lead to LAT phosphorylation. The overall goal of project #1 is to understand the specificity and regulation of protein tyrosine kinases and phosphatases involved In TCR signaling that lead to ITAM and LAT phosphorylation at the membrane surface. We will: 1) define the molecular basis for specificity for Lck and ZAP-70 for ITAMs and LAT, respectively;2) develop an analog sensitive inhibitor system for Lck;3) define the mechanisms that control ITAM phosphorylation by Lck;and, 4) define the mechanisms that control LAT phosphorylation by ZAP-70.

Public Health Relevance

Biochemical signaling events by the T cell receptor (TCR) for antigen controls T cell responses but our understanding of these signaling events Is very rudimentary. These studies aim to understand how the enzymes, kinases and phosphatases, that the TCR controls are regulated on a surface model membrane, as they would be in a cell. These studies could lead to novel therapies of T cells mediated diseases.

Agency
National Institute of Health (NIH)
Institute
National Institute of Allergy and Infectious Diseases (NIAID)
Type
Research Program Projects (P01)
Project #
5P01AI091580-03
Application #
8503585
Study Section
Special Emphasis Panel (ZAI1-RRS-I)
Project Start
2013-07-01
Project End
2016-06-30
Budget Start
2013-07-01
Budget End
2014-06-30
Support Year
3
Fiscal Year
2013
Total Cost
$863,625
Indirect Cost
$128,726
Name
University of California San Francisco
Department
Type
DUNS #
094878337
City
San Francisco
State
CA
Country
United States
Zip Code
94143
Courtney, Adam H; Amacher, Jeanine F; Kadlecek, Theresa A et al. (2017) A Phosphosite within the SH2 Domain of Lck Regulates Its Activation by CD45. Mol Cell 67:498-511.e6
Vercoulen, Yvonne; Kondo, Yasushi; Iwig, Jeffrey S et al. (2017) A Histidine pH sensor regulates activation of the Ras-specific guanine nucleotide exchange factor RasGRP1. Elife 6:
Ashouri, Judith F; Weiss, Arthur (2017) Endogenous Nur77 Is a Specific Indicator of Antigen Receptor Signaling in Human T and B Cells. J Immunol 198:657-668
Ahsan, Nagib; Salomon, Arthur R (2017) Quantitative Phosphoproteomic Analysis of T-Cell Receptor Signaling. Methods Mol Biol 1584:369-382
Myers, Darienne R; Zikherman, Julie; Roose, Jeroen P (2017) Tonic Signals: Why Do Lymphocytes Bother? Trends Immunol 38:844-857
Myers, Darienne R; Lau, Tannia; Markegard, Evan et al. (2017) Tonic LAT-HDAC7 Signals Sustain Nur77 and Irf4 Expression to Tune Naive CD4 T Cells. Cell Rep 19:1558-1571
Bandaru, Pradeep; Shah, Neel H; Bhattacharyya, Moitrayee et al. (2017) Deconstruction of the Ras switching cycle through saturation mutagenesis. Elife 6:
Hsu, Lih-Yun; Cheng, Debra A; Chen, Yiling et al. (2017) Destabilizing the autoinhibitory conformation of Zap70 induces up-regulation of inhibitory receptors and T cell unresponsiveness. J Exp Med 214:833-849
Courtney, Adam H; Lo, Wan-Lin; Weiss, Arthur (2017) TCR Signaling: Mechanisms of Initiation and Propagation. Trends Biochem Sci :
Huang, William Y C; Ditlev, Jonathon A; Chiang, Han-Kuei et al. (2017) Allosteric Modulation of Grb2 Recruitment to the Intrinsically Disordered Scaffold Protein, LAT, by Remote Site Phosphorylation. J Am Chem Soc 139:18009-18015

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