Dual fatty acylation of p59(Fyn) is required for association with the T cell receptor ζ chain through phosphotyrosine-Src homology domain-2 interactions Journal Article
| Authors: | van't Hof, W.; Resh, M. D. |
| Article Title: | Dual fatty acylation of p59(Fyn) is required for association with the T cell receptor ζ chain through phosphotyrosine-Src homology domain-2 interactions |
| Abstract: | The first 10 residues within the Src homology domain (SH)-4 domain of the Src family kinase Fyn are required for binding to the immune receptor tyrosine-based activation motif (ITAM) of T cell receptor (TCR) subunits. Recently, mutation of glycine 2, cysteine 3, and lysines 7 and 9 was shown to block binding of Fyn to TCR ζ chain ITAMs, prompting the designation of these residues as an ITAM recognition motif (Gauen, L.K.T., M.E. Linder, and A.S. Shaw. 1996. J. Cell Biol. 133:1007-1015). Here we show that these residues do not mediate direct interactions with TCR ITAMs, but rather are required for efficient myristoylation and palmitoylation of Fyn. Specifically, coexpression of a K7,9A-Fyn mutant with N- myristoyltransferase restored myristoylation, membrane binding, and association with the cytoplasmic tail of TCR ζ fused to CD8. Conversely, treatment of cells with 2-hydroxymyristate, a myristoylation inhibitor, blocked association of wild type Fyn with ζ. The Fyn NH2 terminus was necessary but not sufficient for interaction with ζ and both Fyn kinase and SH2 domains were required, directing phosphorylation of ζ ITAM tyrosines and binding to ITAM phosphotyrosines. Fyn/ζ interaction was sensitive to octylglucoside and filipin, agents that disrupt membrane rafts. Moreover, a plasma membrane bound, farnesylated Fyn construct, G2A,C3S-FynKRas, was not enriched in the detergent insoluble fraction and did not associate with ζ. We conclude that the Fyn SH4 domain provides the signals for fatty acylation and specific plasma membrane localization, stabilizing the interactions between the Fyn SH2 domain and phosphotyrosines in TCR ζ chain ITAMs. |
| Keywords: | controlled study; protein expression; protein phosphorylation; proto-oncogene proteins; sequence deletion; nonhuman; protein domain; protein localization; cd8 antigen; animal cell; animals; membrane proteins; protein interaction; enzyme activity; transfection; protein tyrosine kinase; cos cells; animalia; t lymphocyte receptor; amino acid sequence; protein processing, post-translational; sequence homology, amino acid; amino terminal sequence; receptors, antigen, t-cell; sequence alignment; recombinant proteins; cell membrane; binding sites; protein-tyrosine kinases; sequence homology; membrane binding; phosphotyrosine; acylation; palmitoylation; proto-oncogene proteins c-fyn; protein kinase p60; myristic acid; myristylation; protein n myristoyltransferase; protein-tyrosine kinase; humans; priority journal; article; src homology domains; receptor/antigen |
| Journal Title: | Journal of Cell Biology |
| Volume: | 145 |
| Issue: | 2 |
| ISSN: | 0021-9525 |
| Publisher: | Rockefeller University Press |
| Date Published: | 1999-04-19 |
| Start Page: | 377 |
| End Page: | 389 |
| Language: | English |
| DOI: | 10.1083/jcb.145.2.377 |
| PUBMED: | 10209031 |
| PROVIDER: | scopus |
| PMCID: | PMC2133112 |
| DOI/URL: | |
| Notes: | Export Date: 18 November 2015 -- Source: Scopus |
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