Tyr¹⁹² Regulates Lymphocyte-Specific Tyrosine Kinase Activity in T Cells

TCR signaling critically depends on the tyrosine kinase Lck (lymphocyte-specific protein tyrosine kinase). Two phosphotyrosines, the activating pTyr³⁹⁴ and the inhibitory pTyr⁵⁰⁵, control Lck activity. Recently, pTyr¹⁹² in the Lck SH2 domain emerged as a third regulator. How pTyr¹⁹² may affect Lck function remains unclear. In this study, we explored the role of Lck Tyr¹⁹² using CRISPR/Cas9-targeted knock-in mutations in the human Jurkat T cell line. Our data reveal that both Lck pTyr³⁹⁴ and pTyr⁵⁰⁵ are controlled by Lck Tyr¹⁹² Lck with a nonphosphorylated SH2 domain (Lck Phe¹⁹²) displayed hyperactivity, possibly by promoting Lck Tyr³⁹⁴ transphosphorylation. Lck Glu¹⁹² mimicking stable Lck pTyr¹⁹² was inhibited by Tyr⁵⁰⁵ hyperphosphorylation. To overcome this effect, we further mutated Tyr⁵⁰⁵ The resulting Lck Glu¹⁹²/Phe⁵⁰⁵ displayed strongly increased amounts of pTyr³⁹⁴ both in resting and activated T cells. Our results suggest that a fundamental role of Lck pTyr¹⁹² may be to protect Lck pTyr³⁹⁴ and/or pTyr⁵⁰⁵ to maintain a pool of already active Lck in resting T cells. This provides an additional mechanism for fine-tuning of Lck as well as T cell activity.