title: Myosin IIA modulates T cell receptor transport and CasL phosphorylation during early immunological synapse formation.
abstract: Activation of T cell receptor (TCR) by antigens occurs in concert with an elaborate multi-scale spatial reorganization of proteins at the immunological synapse, the junction between a T cell and an antigen-presenting cell (APC). The directed movement of molecules, which intrinsically requires physical forces, is known to modulate biochemical signaling. It remains unclear, however, if mechanical forces exert any direct influence on the signaling cascades. We use T cells from AND transgenic mice expressing TCRs specific to the moth cytochrome c 88-103 peptide, and replace the APC with a synthetic supported lipid membrane. Through a series of high spatiotemporal molecular tracking studies in live T cells, we demonstrate that the molecular motor, non-muscle myosin IIA, transiently drives TCR transport during the first one to two minutes of immunological synapse formation. Myosin inhibition reduces calcium influx and colocalization of active ZAP-70 (zeta-chain associated protein kinase 70) with TCR, revealing an influence on signaling activity. More tellingly, its inhibition also significantly reduces phosphorylation of the mechanosensing protein CasL (Crk-associated substrate the lymphocyte type), raising the possibility of a direct mechanical mechanism of signal modulation involving CasL.
authors.affiliation: Howard Hughes Medical Institute, Department of Chemistry, University of California, Berkeley, California, United States of America.
authors.forename: Yan
authors.initials: Y
authors.lastname: Yu
authors.forename: Nicole C
authors.initials: NC
authors.lastname: Fay
authors.forename: Alexander A
authors.initials: AA
authors.lastname: Smoligovets
authors.forename: Hung-Jen
authors.initials: HJ
authors.lastname: Wu
authors.forename: Jay T
authors.initials: JT
authors.lastname: Groves
mesh_terms: D048868:Adaptor Proteins, Signal Transducing / Q000378:metabolism*; D000818:Animals; D001692:Biological Transport / Q000276:immunology; D050719:Crk-Associated Substrate Protein / Q000378:metabolism; D003580:Cytochromes; D054992:Immunological Synapses / Q000378:metabolism*; D051379:Mice; D008822:Mice, Transgenic; D009036:Moths; D024507:Nonmuscle Myosin Type IIA / Q000502:physiology*; D010446:Peptide Fragments; D010766:Phosphorylation; D011948:Receptors, Antigen, T-Cell / Q000378:metabolism*; D015398:Signal Transduction; D013601:T-Lymphocytes
chemical_list: D048868:Adaptor Proteins, Signal Transducing; D050719:Crk-Associated Substrate Protein; D003580:Cytochromes; C505592:NEDD9 protein, mouse; D010446:Peptide Fragments; D011948:Receptors, Antigen, T-Cell; D024507:Nonmuscle Myosin Type IIA