In VivoInteraction of AF-6 with Activated Ras and ZO-1

AF-6 contains two putative Ras-associating domains (RA domains) which are seen in several Ras effectors 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-1in vitro.In this study, we showed t...

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Veröffentlicht in:Biochemical and biophysical research communications 1999-05, Vol.259 (1), p.103-107
Hauptverfasser: Yamamoto, Takaharu, Harada, Naozumi, Kawano, Yoji, Taya, Shinichiro, Kaibuchi, Kozo
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Sprache:eng
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Zusammenfassung:AF-6 contains two putative Ras-associating domains (RA domains) which are seen in several Ras effectors 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-1in 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-fospromoter/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-1in vivo.
ISSN:0006-291X
1090-2104
DOI:10.1006/bbrc.1999.0731