Akt phosphorylation of serine 21 on Pak1 modulates Nck binding and cell migration

被引:144
作者
Zhou, GL
Zhuo, Y
King, CC
Fryer, BH
Bokoch, GM
Field, J [1 ]
机构
[1] Univ Penn, Sch Med, Dept Pharmacol, Philadelphia, PA 19104 USA
[2] Scripps Res Inst, Dept Immunol, La Jolla, CA 92037 USA
[3] Scripps Res Inst, Dept Cell Biol, La Jolla, CA 92037 USA
[4] Univ Calif San Diego, Dept Pharmacol, La Jolla, CA 92093 USA
关键词
D O I
10.1128/MCB.23.22.8058-8069.2003
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 p21-activated protein kinases (Paks) regulate cellular proliferation, differentiation, transformation, and survival through multiple downstream signals. Paks are activated directly by the small GTPases Rac and Cdc42 and several protein kinases including Akt and PDK-1. We found that Akt phosphorylated and modestly activated Pak1 in vitro. The major site phosphorylated by Akt on Pak1 mapped to serine 21, a site originally shown to be weakly autophosphorylated on Pak1 when Cdc42 or Rac activates it. A peptide derived from the region surrounding serine 21 was a substrate for Akt but not Pak1 in vitro, and Akt stimulated serine 21 phosphorylation on the full-length Pak1 much better than Rac did. The adaptor protein Nck binds Pak near serine 21, and its association is regulated by phosphorylation of this site. We found that either treatment of Pak1 in vitro with Akt or coexpression of constitutively active Akt with Pak1 reduced Nck binding to Pak1. In HeLa cells, green fluorescent protein-tagged Pak1 was concentrated at focal adhesions and was released when Akt was cotransfected. A peptide containing the Nck binding site of Pak1 fused to a portion of human immunodeficiency virus Tat to allow it to enter cells was used to test the functional importance of Nck/Pak binding in Akt-stimulated cell migration. This Tat-Nck peptide reduced Akt-stimulated cell migration. Together, these data suggest that Akt modulates the association of Pak with Nck to regulate cell migration.
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页码:8058 / 8069
页数:12
相关论文
共 57 条
[1]   PAK4, a novel effector for Cdc42Hs, is implicated in the reorganization of the actin cytoskeleton and in the formation of filopodia [J].
Abo, A ;
Qu, J ;
Cammarano, MS ;
Dan, CT ;
Fritsch, A ;
Baud, V ;
Belisle, B ;
Minden, A .
EMBO JOURNAL, 1998, 17 (22) :6527-6540
[2]   A novel regulator of p21-activated kinases [J].
Bagrodia, S ;
Taylor, SJ ;
Jordon, KA ;
Van Aelst, L ;
Cerione, RA .
JOURNAL OF BIOLOGICAL CHEMISTRY, 1998, 273 (37) :23633-23636
[3]  
BAGRODIA S, 1995, J BIOL CHEM, V270, P27995
[4]   Pak1 phosphorylation on T212 affects microtubules in cells undergoing mitosis [J].
Banerjee, M ;
Worth, D ;
Prowse, DM ;
Nikolic, M .
CURRENT BIOLOGY, 2002, 12 (14) :1233-1239
[5]   Ras and Rho GTPases: A family reunion [J].
Bar-Sagi, D ;
Hall, A .
CELL, 2000, 103 (02) :227-238
[6]   Interaction of the Nck adapter protein with p21-activated kinase (PAK1) [J].
Bokoch, GM ;
Wang, Y ;
Bohl, BP ;
Sells, MA ;
Quilliam, LA ;
Knaus, UG .
JOURNAL OF BIOLOGICAL CHEMISTRY, 1996, 271 (42) :25746-25749
[7]  
Boyle WJ., 1991, METHOD ENZYMOL, V201, P110
[8]   Human Ste20 homologue hPAK1 links GTPases to the JNK MAP kinase pathway [J].
Brown, JL ;
Stowers, L ;
Baer, M ;
Trejo, J ;
Coughlin, S ;
Chant, J .
CURRENT BIOLOGY, 1996, 6 (05) :598-605
[9]   Akt promotes cell survival by phosphorylating and inhibiting a forkhead transcription factor [J].
Brunet, A ;
Bonni, A ;
Zigmond, MJ ;
Lin, MZ ;
Juo, P ;
Hu, LS ;
Anderson, MJ ;
Arden, KC ;
Blenis, J ;
Greenberg, ME .
CELL, 1999, 96 (06) :857-868
[10]   p21-activated kinase has substrate specificity similar to Acanthamoeba myosin I heavy chain kinase and activates Acanthamoeba myosin I [J].
Brzeska, H ;
Knaus, UG ;
Wang, ZY ;
Bokoch, GM ;
Korn, ED .
PROCEEDINGS OF THE NATIONAL ACADEMY OF SCIENCES OF THE UNITED STATES OF AMERICA, 1997, 94 (04) :1092-1095