| Project/Area Number |
11670442
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| Research Category |
Grant-in-Aid for Scientific Research (C)
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| Allocation Type | Single-year Grants |
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| Section | 一般 |
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| Research Field |
内科学一般
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| Research Institution | University of Tokyo |
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Principal Investigator |
HONDA Zenichire University of Tokyo, Faculty of Medicin Lecturer., 医学部・附属病院, 講師 (70238814)
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| Co-Investigator(Kenkyū-buntansha) |
YOKOMIZO Takehiko University of Tokyo, Bioclieuntry, Assistant., 医学系研究科, 助手 (60302840)
KAZUHIKE Kume University of Tokyo, Bioclieuntry, Assistant., 医学系研究科, 助手 (30251218)
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| Project Period (FY) |
1999 – 2000
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| Project Status |
Completed (Fiscal Year 2000)
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| Budget Amount *help |
¥3,800,000 (Direct Cost: ¥3,800,000)
Fiscal Year 2000: ¥1,500,000 (Direct Cost: ¥1,500,000)
Fiscal Year 1999: ¥2,300,000 (Direct Cost: ¥2,300,000)
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| Keywords | src family kinase / Fc receptors / likid royts / Src family kinases / Fe受容体 / Src型チロシンキナーゼ / 貧食 / raft / Lyn / Fyn / Hck / c-Src |
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| Research Abstract |
Initial biochemical signaling originating from high-affinity immunoglobulin E receptor (FcεRI) has been ascribed to Src family kinases. To understand the mechanisms by which individual kinases drive the signaling, we conducted reconstitution experiments : FcεRI signaling in RBL2H3 cells was first suppressed by a membrane-anchored, gain-of-function C-terminal Src kinase and then reconstructed with Src family kinases whose C-terminal negative regulatory sequence was replaced with a-myc epitope. Those constructs derived from Lyn and Fyn, which are associated with detergent-resistant membranes (DRMs), physically interacted with resting FcεRI and reconstructed clustering-induced signaling that leads to calcium mobilization and ERK1 and-2 activation. c-Src-derived construct, which was excluded from DRMs, failed to interact with FcεRI and to restore the signaling, whereas creation of palmitoylatable Cys3 enabled it to interact with DRMs and with FcεRI and to restore the signaling. Deletion of Src homology 3 (SH3) domain from the Lyn-derived construct did not alter its ability to transduce the series or signaling. Deletion of SH2 domain did not affect its association with DRMs and with FcεRI nor clustering-induced tyrosine phosphorylation of FcεRI β and γ subunits, but it almost abrogated the next step of tyrosine phosphorylation of syk and its recruitment to FcεRI.These findings suggest that Lyn and Fyn could, but c-Src could not, drive FcεRI signaling and that N-terminal palmitoylation and SH2 domain are required in sequence for the initial interaction with FcεRI and for the signal progression to the molecular assembly.
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