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Activator of G protein signaling 3 forms a complex with resistance to inhibitors of cholinesterase-8A without promoting nucleotide exchange on Gαi3

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Abstract

Activator of G protein signaling 3 (AGS3) is a guanine nucleotide dissociation inhibitor (GDI) which stabilizes the Gαi/o subunits as an AGS3/Gαi/o-GDP complex. It has recently been demonstrated in reconstitution experiments that the AGS3/Gαi/o-GDP complex may act as a substrate of resistance to inhibitors of cholinesterase 8A (Ric-8A), a guanine exchange factor (GEF) for heterotrimeric Gα proteins. Since the ability of Ric-8A to activate Gαi/o subunits that are bound to AGS3 in a cellular environment has not been confirmed, we thus examined the effect of Ric-8A on cAMP accumulation in HEK293 cells expressing different forms of AGS3 and Gαi3. Co-immunoprecipitation assays indicate that full-length AGS3 and its N- and C-terminal truncated mutants can interact with Ric-8A in HEK293 cells. Yeast two-hybrid assay further confirmed that Ric-8A can directly bind to AGS3S, a short form of AGS3 which is endogenously expressed in heart. However, Ric-8A failed to facilitate Gαi-induced suppression of adenylyl cyclase, suggesting that it may not serve as a GEF for AGS3/Gαi/o-GDP complex in a cellular environment.

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Acknowledgments

We thank Drs. Yijuang Chern and Marie-Claude Amoureux for the provision of cDNAs. This study was supported by the National Key Basic Research Program of China (2013CB530900), Hong Kong RGC (661807), UGC (T13-607/12R), and the Hong Kong Jockey Club.

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Authors and Affiliations

  1. Division of Life Science and the Biotechnology Research Institute, Hong Kong University of Science and Technology, Clear Water Bay, Kowloon, Hong Kong, China

    Man K. Tse, Christina J. Morris, Mingjie Zhang & Yung H. Wong

  2. State Key Laboratory of Molecular Neuroscience, Hong Kong University of Science and Technology, Clear Water Bay, Kowloon, Hong Kong, China

    Mingjie Zhang & Yung H. Wong

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  1. Man K. Tse
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  2. Christina J. Morris
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Correspondence to Yung H. Wong.

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Tse, M.K., Morris, C.J., Zhang, M. et al. Activator of G protein signaling 3 forms a complex with resistance to inhibitors of cholinesterase-8A without promoting nucleotide exchange on Gαi3 . Mol Cell Biochem 401, 27–38 (2015). https://doi.org/10.1007/s11010-014-2289-7

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