A PLC-gamma 1 Feedback Pathway Regulates Lck Substrate Phosphorylation at the T-Cell Receptor and SLP-76 Complex

摘要

Phospholipase C gamma 1 (PLC-gamma 1) occupies a critically important position in the T-cell signaling pathway. While its functions as a regulator of both Ca2+ signaling and PKC-family kinases are well characterized, PLC-gamma 1's role in the regulation of early T-cell receptor signaling events is incompletely understood. Activation of the T-cell receptor leads to the formation of a signalosome complex between SLP-76, LAT, PLC-gamma 1, Itk, and Vavl. Recent studies have revealed the existence of both positive and negative feedback pathways from SLP-76 to the apical kinase in the pathway, Lck. To determine if PLC-gamma 1 contributes to the regulation of these feedback networks, we performed a quantitative phosphoproteomic analysis of PLC-gamma 1-deficient T cells. These data revealed a previously unappreciated role for PLC-gamma 1 in the positive regulation of Zap-70 and T-cell receptor tyrosine phosphorylation. Conversely, PLC-gamma 1 negatively regulated the phosphorylation of SLP-76-associated proteins, including previously established Lck substrate phosphorylation sites within this complex. While the positive and negative regulatory phosphorylation sites on Lck were largely unchanged, Tyr(192) phosphorylation was elevated in Jgammal. The data supports a model wherein Lck's targeting, but not its kinase activity, is altered by PLC-gamma l, possibly through Lck Tyr(192) phosphorylation and increased association of the kinase with protein scaffolds SLP-76 and TSAd.