The Clk/Sty protein kinase phosphorylates SR splicing factors and regulates their intranuclear distribution

被引:475
作者
Colwill, K
Pawson, T
Andrews, B
Prasad, J
Manley, JL
Bell, JC
Duncan, PI
机构
[1] MT SINAI HOSP, SAMUEL LUNENFELD RES INST, PROGRAMME MOLEC BIOL & CANC, TORONTO, ON M5G 1X5, CANADA
[2] UNIV TORONTO, DEPT MOLEC & MED GENET, TORONTO, ON M5S 1A8, CANADA
[3] COLUMBIA UNIV, DEPT BIOL SCI, NEW YORK, NY 10027 USA
[4] UNIV OTTAWA, OTTAWA REG CANC CTR, CANC RES LABS, OTTAWA, ON K1H 8L6, CANADA
关键词
ASF; protein phosphorylation; RS domains; SF2;
D O I
10.1002/j.1460-2075.1996.tb00357.x
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
Mammalian Clk/Sty is the prototype for a family of dual specificity kinases (termed LAMMER kinases) that have been conserved in evolution, but whose physiological substrates are unknown. In a yeast two-hybrid screen, the Clk/Sty kinase specifically interacted with RNA binding proteins, particularly members of the serine/arginine-rich (SR) family of splicing factors, Clk/Sty itself has an serine/arginine-rich non-catalytic N-terminal region which is important for its association with SR splicing factors. In vitro, Clk/Sty efficiently phosphorylated the SR family member ASF/SF2 on serine residues located within its serine/arginine-rich region (the RS domain). Tryptic phosphopeptide mapping demonstrated that the sites on ASF/SF2 phosphorylated in vitro overlap with those phosphorylated in vivo, Immunofluorescence studies showed that a catalytically inactive form of Clk/Sty co-localized with SR proteins in nuclear speckles, Overexpression of the active Clk/Sty kinase caused a redistribution of SR proteins within the nucleus. These results suggest that the Clk/Sty kinase directly regulates the activity and compartmentalization of SR splicing factors.
引用
收藏
页码:265 / 275
页数:11
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