Cleavage arrest of early frog embryos by the G protein-activated protein kinase PAK I.
PAK I is a member of the PAK (p21-activated protein kinase) family and is activated by Cdc42 (Jakobi, R., Chen, C.-J., Tuazon, P. T., and Traugh, J. A. (1996) J. Biol. Chem. 271, 6206-6211). To examine the effects of PAK I on cleavage arrest, subfemtomole amounts of endogenously active (58 kDa) and inactive (60 kDa) PAK I and a tryptic peptide (37 kDa) containing the active catalytic domain were injected into one blastomere of 2-cell frog embryos. Active PAK I resulted in cleavage arrest in the injected blastomere at mitotic metaphase, whereas the uninjected blastomere progressed through mid- to late cleavage. Injection of other protein kinases at similar concentrations had no effect on cleavage. Endogenous PAK I was highly active in frog oocytes, and antibody to PAK I reacted specifically with protein of 58-60 kDa. PAK I protein was decreased at 60 min post-fertilization, with little or no PAK I protein or activity detectable at 80 min post-fertilization or in 2-cell embryos. At the 4-cell stage PAK I protein increased, but the protein kinase was present primarily as an inactive form. Rac2 and Cdc42, but not Rac 1, were identified in oocytes and throughout early embryo development. Thus, PAK I appears to be a potent cytostatic protein kinase involved in maintaining cells in a non-dividing state. PAK I activity is high in oocytes and appears to be regulated by degradation/synthesis and through autophosphorylation via binding of Cdc42. PAK I may act through regulation of the stress-activated protein kinase signaling pathway and/or by direct regulation of multiple metabolic pathways.
Rooney
R D
RD
Department of Biochemistry, University of California, Riverside, California 92521, USA.
Tuazon
P T
PT
Meek
W E
WE
Carroll
E J
EJ
Hagen
J J
JJ
Gump
E L
EL
Monnig
C A
CA
Lugo
T
T
Traugh
J A
JA
D000818:Animals; D001001:Anura / Q000196:embryology*; D001757:Blastomeres / Q000201:enzymology / Q000378:metabolism; D018797:Cell Cycle Proteins / Q000378:metabolism; D002455:Cell Division; D002970:Cleavage Stage, Ovum / Q000166:cytology*; D019204:GTP-Binding Proteins / Q000378:metabolism*; D009865:Oocytes / Q000201:enzymology; D011494:Protein Kinases / Q000378:metabolism*; D020744:rac GTP-Binding Proteins
D018797:Cell Cycle Proteins; D011494:Protein Kinases; C036447:protease activated kinase I; D019204:GTP-Binding Proteins; D020744:rac GTP-Binding Proteins