TY - JOUR
T1 - In vivo interaction of AF-6 with activated Ras and ZO-1
AU - Yamamoto, Takaharu
AU - Harada, Naozumi
AU - Kawano, Yoji
AU - Taya, Shinichiro
AU - Kaibuchi, Kozo
N1 - Funding Information:
This study is supported by grants-in-aid for Scientific Research from the Ministry of Education, Science, and Culture, Japan, by a grant from Japan Society of the Promotion of Science Research for the Future, and by a grant from Kirin Brewery Company Limited. T. Yamamoto and S. Taya are research fellows of Japan Society for the Promotion of Science.
PY - 1999/5/27
Y1 - 1999/5/27
N2 - AF-6 contains two putative Ras-associating domains (RA domains) which are seen in several Ras effecters such as RalGDS and RIN1. We previously showed that an AF-6 fragment containing the amino-terminal (N-terminal) RA domain directly binds to activated Ras and ZO-1 in vitro. In this study, we showed that a single amino acid mutation in the N-terminal RA domain of AF-6 abolished the interaction of AF-6 with activated Ras and that the sites of this critical amino acid residue were similar to those for Raf-1 and RalGDS. The overexpression of the N-terminal RA domain of AF-6 inhibited the Ras-dependent c-fos promoter/enhancer stimulation in NIH3T3 cells. Endogenous AF-6 was coimmunoprecipitated with activated Ras from Rat1 cells expressing activated Ras. Moreover, we showed that AF-6 was coimmunoprecipitated with ZO-1 from Rat1 cells. Taken together, these results indicate that the Ras-interacting region on AF-6 is structurally similar to that on Raf-1 and on RalGDS and that AF-6 interacts with activated Ras and ZO-1 in vivo.
AB - AF-6 contains two putative Ras-associating domains (RA domains) which are seen in several Ras effecters such as RalGDS and RIN1. We previously showed that an AF-6 fragment containing the amino-terminal (N-terminal) RA domain directly binds to activated Ras and ZO-1 in vitro. In this study, we showed that a single amino acid mutation in the N-terminal RA domain of AF-6 abolished the interaction of AF-6 with activated Ras and that the sites of this critical amino acid residue were similar to those for Raf-1 and RalGDS. The overexpression of the N-terminal RA domain of AF-6 inhibited the Ras-dependent c-fos promoter/enhancer stimulation in NIH3T3 cells. Endogenous AF-6 was coimmunoprecipitated with activated Ras from Rat1 cells expressing activated Ras. Moreover, we showed that AF-6 was coimmunoprecipitated with ZO-1 from Rat1 cells. Taken together, these results indicate that the Ras-interacting region on AF-6 is structurally similar to that on Raf-1 and on RalGDS and that AF-6 interacts with activated Ras and ZO-1 in vivo.
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U2 - 10.1006/bbrc.1999.0731
DO - 10.1006/bbrc.1999.0731
M3 - Article
C2 - 10334923
AN - SCOPUS:0033609367
SN - 0006-291X
VL - 259
SP - 103
EP - 107
JO - Biochemical and Biophysical Research Communications
JF - Biochemical and Biophysical Research Communications
IS - 1
ER -