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J Biol Chem, Vol. 273, Issue 52, 34663-34666, December 25, 1998
From the Division of Signal Transduction, Nara Institute of Science
and Technology, 8916-5 Takayama, Ikoma 630-0101, Japan
Rho-associated kinase (Rho-kinase), which is
activated by the small GTPase Rho, phosphorylates moesin at
Thr558 in vitro. Here, using a site- and
phosphorylation state-specific antibody, we found that the expression
of dominant active RhoA in COS7 cells induced moesin phosphorylation
and the formation of microvilli-like structures at apical membranes
where the Thr558-phosphorylated moesin accumulated, whereas
the expression of dominant negative Rho-kinase inhibited both of these
processes. The expression of dominant active Rho-kinase also induced
moesin phosphorylation. When COS7 cells expressing moesin or
moesinT558A (substitution of Thr by Ala) were cultured
under serum-depleted conditions, there were few microvilli-like
structures, whereas microvilli-like structures remained in the cells
expressing moesinT558D (substitution of Thr by Asp). The
expression of moesinT558A inhibited the dominant active
RhoA-induced formation of microvilli-like structures. These results
indicate that Rho-kinase regulates moesin phosphorylation downstream of
Rho in vivo and that the phosphorylation of moesin by
Rho-kinase plays a crucial role in the formation of microvilli-like structures.
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