title: Structures of the APC-ARM domain in complexes with discrete Amer1/WTX fragments reveal that it uses a consensus mode to recognize its binding partners.
abstract: The tumor suppressor APC employs its conserved armadillo repeat (ARM) domain to recognize many of its binding partners, including Amer1/WTX, which is mutated in Wilms' tumor and bone overgrowth syndrome. The APC-Amer1 complex has important roles in regulating Wnt signaling and cell adhesion. Three sites A1, A2, and A3 of Amer1 have been reported to mediate its interaction with APC-ARM. In this study, crystal structures of APC-ARM in complexes with Amer1-A1, -A2, and -A4, which is newly identified in this work, were determined. Combined with our GST pull-down, yeast two-hybrid, and isothermal titration calorimetry (ITC) assay results using mutants of APC and Amer1 interface residues, our structures demonstrate that Amer1-A1, -A2, and -A4, as well as other APC-binding proteins such as Asef and Sam68, all employ a common recognition pattern to associate with APC-ARM. In contrast, Amer1-A3 binds to the C-terminal side of APC-ARM through a bipartite interaction mode. Composite mutations on either APC or Amer1 disrupting all four interfaces abrogated their association in cultured cells and impaired the membrane recruitment of APC by Amer1. Our study thus comprehensively elucidated the recognition mechanism between APC and Amer1, and revealed a consensus recognition sequence employed by various APC-ARM binding partners.
authors.affiliation: School of Life Sciences and Biotechnology, State Key Laboratory of Microbial Metabolism, Shanghai Jiao Tong University , Shanghai, China.
authors.forename: Zhenyi
authors.initials: Z
authors.lastname: Zhang
authors.affiliation: Nikolaus-Fiebiger-Center for Molecular Medicine, University Erlangen-Nuremberg , Erlangen, Germany.
authors.forename: Senem
authors.initials: S
authors.lastname: Akyildiz
authors.affiliation: School of Life Sciences and Biotechnology, State Key Laboratory of Microbial Metabolism, Shanghai Jiao Tong University , Shanghai, China.
authors.forename: Yafei
authors.initials: Y
authors.lastname: Xiao
authors.affiliation: School of Life Sciences and Biotechnology, State Key Laboratory of Microbial Metabolism, Shanghai Jiao Tong University , Shanghai, China.
authors.forename: Zhongchao
authors.initials: Z
authors.lastname: Gai
authors.affiliation: School of Life Sciences and Biotechnology, State Key Laboratory of Microbial Metabolism, Shanghai Jiao Tong University , Shanghai, China.
authors.forename: Ying
authors.initials: Y
authors.lastname: An
authors.affiliation: Nikolaus-Fiebiger-Center for Molecular Medicine, University Erlangen-Nuremberg , Erlangen, Germany.
authors.forename: Jürgen
authors.initials: J
authors.lastname: Behrens
authors.affiliation: School of Life Sciences and Biotechnology, State Key Laboratory of Microbial Metabolism, Shanghai Jiao Tong University , Shanghai, China.
authors.forename: Geng
authors.initials: G
authors.lastname: Wu
keywords: APC; ARM domain; Amer1; WTX; consensus recognition mode; crystal structure