Distinct structural and catalytic roles for Zap70 in formation of the immunological synapse in CTL

• Type of Material: Article
• Authors: Misty R. Jenkins, University of Cambridge; Jane C. Stinchcombe, University of Cambridge; Byron Au-Yeung, Emory University; Yukako Asano, University of Cambridge; Alex T. Ritter, University of Cambridge; Arthur Weiss, University of California San Francisco; Gillian M Griffiths, University of Cambridge
• Language: English
• Date: 2014-03-04
• Publisher: eLife Sciences Publications
• Publication Version: Final Published Version
• Copyright Statement: © Jenkins et al.
• License: Creative Commons Attribution 3.0 Unported
• Final Published Version (URL): http://dx.doi.org/10.7554/eLife.01310
• Title of Journal or Parent Work: eLife
• ISSN: 2050-084X
• Volume: 3
• Issue: 3
• Start Page: e01310
• End Page: e01310
• Grant/Funding Information: Australian National Health and Medical Research Council (NHMRC) Biomedical Fellowship 567082 to Misty R Jenkins.; Arthritis Foundation 5476 to Byron B Au-Yeung.; NIAMS, National Institutes of Health RC2AR058947 to Arthur Weiss.; This paper was supported by the following grants:; Howard Hughes Medical Institute to Arthur Weiss.; Wellcome Trust 075880, 100140 to Gillian M Griffiths.
• Keywords: GRANULE POLARIZATION; ACTIN POLYMERIZATION; Biology; Life Sciences & Biomedicine; TYROSINE PHOSPHORYLATION; RETROGRADE FLOW; SECRETORY GRANULES; Science & Technology; ZAP-70 KINASE; MEDIATED CYTOTOXICITY; NATURAL-KILLER-CELLS; T-CELL-RECEPTOR; Life Sciences & Biomedicine - Other Topics; SEVERE COMBINED IMMUNODEFICIENCY
• Research Categories: Health Sciences, Immunology; Biology, Microbiology

Abstract

T cell receptor (TCR) activation leads to a dramatic reorganisation of both membranes and receptors as the immunological synapse forms. Using a genetic model to rapidly inhibit Zap70 catalytic activity we examined synapse formation between cytotoxic T lymphocytes and their targets. In the absence of Zap70 catalytic activity Vav-1 activation occurs and synapse formation is arrested at a stage with actin and integrin rich interdigitations forming the interface between the two cells. The membranes at the synapse are unable to flatten to provide extended contact, and Lck does not cluster to form the central supramolecular activation cluster (cSMAC). Centrosome polarisation is initiated but aborts before reaching the synapse and the granules do not polarise. Our findings reveal distinct roles for Zap70 as a structural protein regulating integrin-mediated control of actin vs its catalytic activity that regulates TCR-mediated control of actin and membrane remodelling during formation of the immunological synapse.