Phospho‐tyrosine dependent protein–protein interaction network

Molecular Systems Biology

Arndt Grossmann¹  Nouhad Benlasfer¹  Petra Birth¹  Anna Hegele¹  Franziska Wachsmuth¹  Luise Apelt¹ 

[1] Otto-Warburg Laboratory, Max-Planck Institute for Molecular Genetics (MPIMG), Berlin, Germany

关键词: cancer signaling;    network biology;    post‐translational protein modification;    yeast two‐hybrid;   

DOI  :  10.15252/msb.20145968

来源: Wiley

【 摘 要 】

Abstract

Post-translational protein modifications, such as tyrosine phosphorylation, regulate protein–protein interactions (PPIs) critical for signal processing and cellular phenotypes. We extended an established yeast two-hybrid system employing human protein kinases for the analyses of phospho-tyrosine (pY)-dependent PPIs in a direct experimental, large-scale approach. We identified 292 mostly novel pY-dependent PPIs which showed high specificity with respect to kinases and interacting proteins and validated a large fraction in co-immunoprecipitation experiments from mammalian cells. About one-sixth of the interactions are mediated by known linear sequence binding motifs while the majority of pY-PPIs are mediated by other linear epitopes or governed by alternative recognition modes. Network analysis revealed that pY-mediated recognition events are tied to a highly connected protein module dedicated to signaling and cell growth pathways related to cancer. Using binding assays, protein complementation and phenotypic readouts to characterize the pY-dependent interactions of TSPAN2 (tetraspanin 2) and GRB2 or PIK3R3 (p55γ), we exemplarily provide evidence that the two pY-dependent PPIs dictate cellular cancer phenotypes.

Synopsis

A modified yeast two-hybrid approach employed on a large scale generates a network of 292 human phospho-tyrosine (pY)-dependent protein–protein interactions. Conditional interactions are validated, and pY-dependent interaction specificity and network features are assessed.

• A pY-dependent protein interaction data set is generated using a modified yeast two-hybrid approach.
• Network analyses assess the extent of known linear motif-based pY recognition, pointing toward the importance of context for interaction specificity, and reveal a highly connected pY-recognition module in the human proteome.
• A large fraction of PPIs is validated by co-immunoprecipitation with good success rate.
• pY-dependent TSPAN2 interactions are related to cancer phenotypes.

【 授权许可】

CC BY   
© 2015 The Authors. Published under the terms of the CC BY 4.0 license

Creative Commons Attribution 4.0 License, which permits use, distribution and reproduction in any medium, provided the original work is properly cited.