SU6656, a selective Src family kinase inhibitor, used to probe growth factor signaling

被引:519
作者
Blake, RA [1 ]
Broome, MA [1 ]
Liu, XD [1 ]
Wu, JM [1 ]
Gishizky, M [1 ]
Sun, L [1 ]
Courtneidge, SA [1 ]
机构
[1] SUGEN Inc, S San Francisco, CA 94080 USA
关键词
D O I
10.1128/MCB.20.23.9018-9027.2000
中图分类号
Q5 [生物化学]; Q7 [分子生物学];
学科分类号
071010 ; 081704 ;
摘要
The use of small-molecule inhibitors to study molecular components of cellular signal transduction pathways provides a means of analysis complementary to currently used techniques, such as antisense, dominant-negative (interfering) mutants and constitutively activated mutants. We have identified and characterized a small-molecule inhibitor, SU6656, which exhibits selective for Src and other members of the Src family. A related inhibitor, SU6657, inhibits many kinases, including Src and the platelet-derived growth factor (PDGF) receptor. The nse of SU6656 confirmed our previous findings that Src family kinases are required for both Myc induction and DNA synthesis in response to PDGF stimulation of MDI 3T3 fibroblasts. By comparing PDGF stimulated tyrosine phosphorylation events in untreated and SU6656-treated cells, we found that some substrates (for example, c-Cbl, and protein kinase C delta) were Src family substrates whereas others (for example, phospholipase C-gamma) were not. One protein, the adaptor Shc, was a substrate for both Src family kinases ton tyrosines 239 and 240) and a distinct tyrosine kinase ton tyrosine 317, which is perhaps phosphorylated by the PDGF receptor itself). Microinjection experiments demonstrated that a Shc molecule carrying mutations of tyrosines 239 and 240, in conjunction with an SH2 domain mutation, interfered with PDGF-stimulated DNA synthesis. Deletion of the phosphotyrosine-binding domain also inhibited synthesis. These inhibitions were overcome by heterologous expression of Myc, supporting the hypothesis that Shc functions in the Src pathway. SU6656 should prove a useful additional tool for further dissecting the role of Src kinases in this and other signal transduction pathways.
引用
收藏
页码:9018 / 9027
页数:10
相关论文
共 35 条
[1]   MYC BUT NOT FOS RESCUE OF PDGF SIGNALING BLOCK CAUSED BY KINASE INACTIVE SRC [J].
BARONE, MV ;
COURTNEIDGE, S .
NATURE, 1995, 378 (6556) :509-512
[2]  
Blake RA, 1999, CELL GROWTH DIFFER, V10, P231
[3]   The proto-oncogene c-Cbl is a negative regulator of DNA synthesis initiated by both receptor and cytoplasmic tyrosine kinases [J].
Broome, MA ;
Galisteo, ML ;
Schlessinger, J ;
Courtneidge, SA .
ONCOGENE, 1999, 18 (18) :2908-2912
[4]   No requirement for Src family kinases for PDGF signaling in fibroblasts expressing SV40 large T antigen [J].
Broome, MA ;
Courtneidge, SA .
ONCOGENE, 2000, 19 (24) :2867-2869
[5]   Signal-transduction cascades as targets for therapeutic intervention by natural products [J].
Cardenas, ME ;
Sanfridson, A ;
Cutler, NS ;
Heitman, J .
TRENDS IN BIOTECHNOLOGY, 1998, 16 (10) :427-433
[6]   Activation of Src family members is not required for the platelet-derived growth factor β receptor to initiate mitogenesis [J].
DeMali, KA ;
Kazlauskas, A .
MOLECULAR AND CELLULAR BIOLOGY, 1998, 18 (04) :2014-2022
[7]  
Goekjian PG, 1999, CURR MED CHEM, V6, P877
[8]   Tyrosine phosphorylation sites at amino acids 239 and 240 of Shc are involved in epidermal growth factor-induced mitogenic signaling that is distinct from Ras/mitogen-activated protein kinase activation [J].
Gotoh, N ;
Toyoda, M ;
Shibuya, M .
MOLECULAR AND CELLULAR BIOLOGY, 1997, 17 (04) :1824-1831
[9]   Signal transduction via platelet-derived growth factor receptors [J].
Heldin, CH ;
Östman, A ;
Rönnstrand, L .
BIOCHIMICA ET BIOPHYSICA ACTA-REVIEWS ON CANCER, 1998, 1378 (01) :F79-F113
[10]  
Hill RJ, 1996, CELL GROWTH DIFFER, V7, P1125