共查询到20条相似文献,搜索用时 31 毫秒
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G L Shen-Ong R M Skurla Jr J D Owens J F Mushinski 《Molecular and cellular biology》1990,10(6):2715-2722
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Multiple novel transcription initiation sites for NRG1 总被引:5,自引:0,他引:5
Steinthorsdottir V Stefansson H Ghosh S Birgisdottir B Bjornsdottir S Fasquel AC Olafsson O Stefansson K Gulcher JR 《Gene》2004,342(1):97-105
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Korschineck I Ziegler S Breuss J Lang I Lorenz M Kaun C Ambros PF Binder BR 《The Journal of biological chemistry》2001,276(16):13192-13197
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Zamora-Leon SP Lee G Davies P Shafit-Zagardo B 《The Journal of biological chemistry》2001,276(43):39950-39958
Microtubule-associated protein 2 (MAP-2) isoforms are developmentally expressed in the nervous system and contain a number of functional domains. Adjacent to the first repeat of the microtubule-binding domain is an RTPPKSP motif for binding SH3 domains. To identify SH3-containing proteins that interact with MAP-2, transfections, filter overlay assays, glutathione S-transferase (GST)-mediated binding assays, co-immunoprecipitations and enzyme-linked immunosorbent assays were performed. Transfections of MAP-2a, MAP-2b, and MAP-2c constructs into COS7 cells, followed by incubation of the cell lysates with SH3-GST fusion proteins, determined that the strongest interaction was between MAP-2c and the non-receptor tyrosine kinase Fyn; however, MAP-2b and MAP-2c also bound to Grb2. Co-immunoprecipitation of Fyn and MAP-2c from human fetal homogenates confirmed the interaction in vivo. MAP-2 synthetic peptides spanning the RTPPKSP motif bound to Fyn, and the interaction was regulated by phosphorylation. Co-transfections with MAP-2c and the extracellular signal-regulated kinase 2 (ERK2) demonstrated that MAP-2c is threonine/serine-phosphorylated on its RTPPKSP motif and that threonine phosphorylation abolished the MAP-2c/Fyn binding. Kinase assays and co-transfection of MAP-2c and Fyn confirmed that Fyn tyrosine kinase phosphorylates MAP-2c. Thus, the activation of signaling pathways may regulate cytoskeletal dynamics by altering the state of phosphorylation of MAP-2 by both ERK2 and Fyn kinase. 相似文献