TY - JOUR
T1 - Regulation of cross-linking of actin filament by IQGAP1, a target for Cdc42
AU - Fukataf, Masaki
AU - Kuroda, Shinya
AU - Fujii, Katsuhiko
AU - Nakamura, Tomoko
AU - Shoji, Ikuo
AU - Matsuura, Yoshiharu
AU - Okawa, Katsuya
AU - Iwamatsu, Akihiro
AU - Kikuchi, Akira
AU - Kaibuchi, Kozo
PY - 1997/11/21
Y1 - 1997/11/21
N2 - We have previously shown that IQGAP1, a recently identified target for Cdc42 and Rac1 small GTPases, showed a distribution similar to that of cortical actin cytoskeleton at the membrane ruffling area induced by insulin and Rac1(val12) (Kuroda, S., Fukata, M., Kobayashi, K., Nakafuku, M., Nomura, N., Iwamatsu, A., and Kaibuchi, K. (1996) J. Biol. Chem. 271, 23363-23367). Here we identified an IQGAP1-interacting molecule with molecular mass of 43 kDa (p43) from bovine brain cytosol, using glutathione S-transferase (GST)- IQGAP1 affinity column chromatography. The amine acid sequencing of the protein revealed that p43 was identical to β- and γ-actin. IQGAP1 was cosedimentated with filamentous actin (F-actin). The amino-terminal domain (amino acids 1-216) of IQGAP1 was responsible for the interaction with F- actin. Falling ball viscometry assay revealed that IQGAP1 cross-linked the F- actin. This IQGAP1 activity was further enhanced by guanosine 5'-(3-0- thio)triphosphate (GTPγS)-GST-Cdc42 but not by GDP-GST-Cdc42. The gel filtration analysis of IQGAP1 revealed that IQGAP1 appeared as oligomers and that GTPγS-GST-Cdc42 but not GDP-GST-Cdc42 enhanced the oligomerization of IQGAP1. These results strongly suggest that IQGAP1, acting downstream of Cdc42, can cross-link the actin filament through its oligomerization.
AB - We have previously shown that IQGAP1, a recently identified target for Cdc42 and Rac1 small GTPases, showed a distribution similar to that of cortical actin cytoskeleton at the membrane ruffling area induced by insulin and Rac1(val12) (Kuroda, S., Fukata, M., Kobayashi, K., Nakafuku, M., Nomura, N., Iwamatsu, A., and Kaibuchi, K. (1996) J. Biol. Chem. 271, 23363-23367). Here we identified an IQGAP1-interacting molecule with molecular mass of 43 kDa (p43) from bovine brain cytosol, using glutathione S-transferase (GST)- IQGAP1 affinity column chromatography. The amine acid sequencing of the protein revealed that p43 was identical to β- and γ-actin. IQGAP1 was cosedimentated with filamentous actin (F-actin). The amino-terminal domain (amino acids 1-216) of IQGAP1 was responsible for the interaction with F- actin. Falling ball viscometry assay revealed that IQGAP1 cross-linked the F- actin. This IQGAP1 activity was further enhanced by guanosine 5'-(3-0- thio)triphosphate (GTPγS)-GST-Cdc42 but not by GDP-GST-Cdc42. The gel filtration analysis of IQGAP1 revealed that IQGAP1 appeared as oligomers and that GTPγS-GST-Cdc42 but not GDP-GST-Cdc42 enhanced the oligomerization of IQGAP1. These results strongly suggest that IQGAP1, acting downstream of Cdc42, can cross-link the actin filament through its oligomerization.
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U2 - 10.1074/jbc.272.47.29579
DO - 10.1074/jbc.272.47.29579
M3 - Article
C2 - 9368021
AN - SCOPUS:0030697431
SN - 0021-9258
VL - 272
SP - 29579
EP - 29583
JO - Journal of Biological Chemistry
JF - Journal of Biological Chemistry
IS - 47
ER -