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Blood, 1 July 2004, Vol. 104, No. 1, pp. 207-214.
Prepublished online as a Blood First Edition Paper on March 9, 2004; DOI 10.1182/blood-2003-08-2769.
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Submitted August 12, 2003
Accepted March 2, 2004
NTAL phosphorylation is a pivotal link between the signaling cascades leading to human mast cell degranulation following Kit activation and Fc RI aggregation
Christine Tkaczyk, Vaclav Horejsi, Shoko Iwaki, Petr Draber, Lawrence E Samelson, Anne B Satterthwaite, Dong-Ho Nahm, Dean D Metcalfe, and Alasdair M Gilfillan*
Laboratory of Allergic Diseases, National Institute of Allergy and Infectious Diseases, National Institutes of Health, Bethesda, MD, USA
Institute of Molecular Genetics, Academy of Sciences of the Czech Republic, Prague, Czech Republic
Center for Cellular and Molecular Biology, Center for Cancer Research, National Cancer Institute, National Institutes of Health, Bethesda, MD, USA
Department of Internal Medicine, University of Texas Southwestern Medical Center, Dallas, TX, USA
* Corresponding author; email: agilfillan{at}niaid.nih.gov.
Fc RI aggregation on the surface of mast cells results in degranulation, a response which is potentiated by binding of stem cell factor (SCF) to its receptor Kit. We observed that one of the major initial signaling events associated with FcRI-mediated activation of human mast cells (HuMCs) is the rapid tyrosine phosphorylation of a protein of 25-30 kD. The phosphorylation of this protein was also observed in response to SCF. This protein was identified as Non T cell Activation Linker (NTAL), an adaptor molecule similar to LAT. Unlike the FcRI response, SCF induced NTAL phosphorylation in the absence of detectable LAT phosphorylation. When SCF and antigen were added concurrently, there was a marked synergistic effect on NTAL phosphorylation, however, SCF did not enhance the phosphorylation of LAT induced by FcRI aggregation. FcRI- and SCF-mediated NTAL phosphorylation appear to be differentially regulated by Src kinases and / or Kit kinase respectively. Diminution of NTAL expression by silencing RNA oligonucleotides in HuMCs resulted in a reduction of both Kit and FcRI-mediated degranulation. NTAL, thus, appears to be an important link between the signaling pathways which are initiated by these receptors culminating in mast cell degranulation.
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