Cdc42 is required for PIP(2)-induced actin polymerization and early development but not for cell viability.
BACKGROUND
Cdc42 and other Rho GTPases are conserved from yeast to humans and are thought to regulate multiple cellular functions by inducing coordinated changes in actin reorganization and by activating signaling pathways leading to specific gene expression. Direct evidence implicating upstream signals and components that regulate Cdc42 activity or for required roles of Cdc42 in activation of downstream protein kinase signaling cascades is minimal, however. Also, whereas genetic analyses have shown that Cdc42 is essential for cell viability in yeast, its potential roles in the growth and development of mammalian cells have not been directly assessed.


RESULTS
To elucidate potential functions of Cdc42 mammalian cells, we used gene-targeted mutation to inactivate Cdc42 in mouse embryonic stem (ES) cells and in the mouse germline. Surprisingly, Cdc42-deficient ES cells exhibited normal proliferation and phosphorylation of mitogen- and stress-activated protein kinases. Yet Cdc42 deficiency caused very early embryonic lethality in mice and led to aberrant actin cytoskeletal organization in ES cells. Moreover, extracts from Cdc42-deficient cells failed to support phosphatidylinositol 4,5-bisphosphate (PIP(2))-induced actin polymerization.


CONCLUSIONS
Our studies clearly demonstrate that Cdc42 mediates PIP(2)-induced actin assembly, and document a critical and unique role for Cdc42 in this process. Moreover, we conclude that, unexpectedly, Cdc42 is not necessary for viability or proliferation of mammalian early embryonic cells. Cdc42 is, however, absolutely required for early mammalian development.
Chen
F
F
Departments of Genetics, The Center for Blood Research, Harvard Medical School, Boston, MA 02115, USA.
Ma
L
L
Parrini
M C
MC
Mao
X
X
Lopez
M
M
Wu
C
C
Marks
P W
PW
Davidson
L
L
Kwiatkowski
D J
DJ
Kirchhausen
T
T
Orkin
S H
SH
Rosen
F S
FS
Mayer
B J
BJ
Kirschner
M W
MW
Alt
F W
FW
D000199:Actins / Q000187:drug effects* / Q000378:metabolism; D000818:Animals; D016923:Cell Death; D002455:Cell Division; D002460:Cell Line; D002470:Cell Survival; D003599:Cytoskeleton / Q000187:drug effects / Q000378:metabolism; D004622:Embryo, Mammalian / Q000166:cytology / Q000502:physiology*; D004789:Enzyme Activation; D051379:Mice; D018345:Mice, Knockout; D020928:Mitogen-Activated Protein Kinases / Q000378:metabolism; D019269:Phosphatidylinositol 4,5-Diphosphate / Q000494:pharmacology*; D020764:cdc42 GTP-Binding Protein / Q000172:deficiency / Q000235:genetics / Q000378:metabolism*
D000199:Actins; D019269:Phosphatidylinositol 4,5-Diphosphate; D020928:Mitogen-Activated Protein Kinases; D020764:cdc42 GTP-Binding Protein