Phosphorylation and dimerization regulate nucleocytoplasmic shuttling of mammalian STE20-like kinase (MST)

被引:72
作者
Lee, KK [1 ]
Yonehara, S
机构
[1] Kyoto Univ, Inst Virus Res, Sakyo Ku, Kyoto 6068507, Japan
[2] Kyoto Univ, Grad Sch Biostudies, Sakyo Ku, Kyoto 6068507, Japan
关键词
D O I
10.1074/jbc.M108138200
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 STE20-like kinase (MST) is a member of the yeast STE20-related kinase family and proteolytically activated by caspase during apoptosis. However, its other cellular functions are not known, including its activation mechanism, substrate(s), and subcellular localization. In this report, using anti-MST monoclonal antibodies, we clearly show that endogenous MST is localized in cytoplasm in a leptomycin B-dependent manner. Analyses with serial deletions and point mutations show that MST has two functional nuclear export signals and, unexpectedly, another localization motif for nuclear import. When cells are treated with leptomycin, monomeric MST is accumulated more rapidly in the nucleus than dimeric MST, indicating that dimerization contributes to the cytoplasmic retention of MST. Okadaic acid, an inhibitor of phosphatase 2A, induces activation of MST and translocation into the nucleus. Using phosphopeptide-specific antibody, we directly show that okadaic acid induces phosphorylation in the activation loop of MST, and, once phosphorylated, MST is rapidly translocated to the nucleus. However, kinase-deficient MST does not enter the nucleus, indicating that phosphorylation and activation is required for okadaic acid-induced nuclear translocation. In apoptotic cells, the activation of MST does not require phosphorylation in the activation loop and occurs through the release of C-terminal regulatory domain by caspase-dependent cleavage. Kinase-deficient MST functions dominant-negatively and represses okadaic acid-induced morphological change indicating that MST plays a role in okadaic acid-induced cellular shrinkage. Our identification of cytoplasmic and nuclear localization motifs and phosphorylation-dependent translocation of MST suggests that regulation of localization is important to the biological function of MST, including its effects on cellular morphology.
引用
收藏
页码:12351 / 12358
页数:8
相关论文
共 42 条
[1]   Identification of the thyroid transcription factor-1 as a target for rat MST2 kinase [J].
Aurisicchio, L ;
Di Lauro, R ;
Zannini, M .
JOURNAL OF BIOLOGICAL CHEMISTRY, 1998, 273 (03) :1477-1482
[2]   PAK to the future [J].
Bagrodia, S ;
Cerione, RA .
TRENDS IN CELL BIOLOGY, 1999, 9 (09) :350-355
[3]   Nuclear export of the stress-activated protein kinase p38 mediated by its substrate MAPKAP kinase-2 [J].
Ben-Levy, R ;
Hooper, S ;
Wilson, R ;
Paterson, HF ;
Marshall, CJ .
CURRENT BIOLOGY, 1998, 8 (19) :1049-1057
[4]   Caspase-mediated cleavage and functional changes of hematopoietic progenitor kinase 1 (HPK1) [J].
Chen, YR ;
Meyer, CF ;
Ahmed, B ;
Yao, ZB ;
Tan, TH .
ONCOGENE, 1999, 18 (51) :7370-7377
[5]   The Ste20-like protein kinase, Mst1, dimerizes and contains an inhibitory domain [J].
Creasy, CL ;
Ambrose, DM ;
Chernoff, J .
JOURNAL OF BIOLOGICAL CHEMISTRY, 1996, 271 (35) :21049-21053
[6]   CLONING AND CHARACTERIZATION OF A HUMAN PROTEIN-KINASE WITH HOMOLOGY TO STE20 [J].
CREASY, CL ;
CHERNOFF, J .
JOURNAL OF BIOLOGICAL CHEMISTRY, 1995, 270 (37) :21695-21700
[7]   p21-activated protein kinase: a crucial component of morphological signaling? [J].
Daniels, RH ;
Bokoch, GM .
TRENDS IN BIOCHEMICAL SCIENCES, 1999, 24 (09) :350-355
[8]   Leptomycin B-sensitive nuclear export of MAPKAP kinase 2 is regulated by phosphorylation [J].
Engel, K ;
Kotlyarov, A ;
Gaestel, M .
EMBO JOURNAL, 1998, 17 (12) :3363-3371
[9]   ROLE OF NUCLEAR TRAFFICKING IN REGULATING CELLULAR-ACTIVITY [J].
FELDHERR, CM ;
AKIN, D .
INTERNATIONAL REVIEW OF CYTOLOGY - A SURVEY OF CELL BIOLOGY, VOL 151, 1994, 151 :183-228
[10]   INTERLEUKIN-1 ACTIVATES A NOVEL PROTEIN-KINASE CASCADE THAT RESULTS IN THE PHOSPHORYLATION OF HSP27 [J].
FRESHNEY, NW ;
RAWLINSON, L ;
GUESDON, F ;
JONES, E ;
COWLEY, S ;
HSUAN, J ;
SAKLATVALA, J .
CELL, 1994, 78 (06) :1039-1049