title: Comprehensive Characterization of Minichromosome Maintenance Complex (MCM) Protein Interactions Using Affinity and Proximity Purifications Coupled to Mass Spectrometry.
abstract: The extensive identification of protein-protein interactions under different conditions is an important challenge to understand the cellular functions of proteins. Here we use and compare different approaches including affinity purification and purification by proximity coupled to mass spectrometry to identify protein complexes. We explore the complete interactome of the minichromosome maintenance (MCM) complex by using both approaches for all of the different MCM proteins. Overall, our analysis identified unique and shared interaction partners and proteins enriched for distinct biological processes including DNA replication, DNA repair, and cell cycle regulation. Furthermore, we mapped the changes in protein interactions of the MCM complex in response to DNA damage, identifying a new role for this complex in DNA repair. In summary, we demonstrate the complementarity of these approaches for the characterization of protein interactions within the MCM complex.
authors: Department of Anatomy and Cell Biology, Université de Sherbrooke , 3201 Jean-Mignault, Sherbrooke, Québec J1E 4K8, Canada.
authors: Marie-Line
authors: ML
authors: Dubois
authors: Department of Anatomy and Cell Biology, Université de Sherbrooke , 3201 Jean-Mignault, Sherbrooke, Québec J1E 4K8, Canada.
authors: Charlotte
authors: C
authors: Bastin
authors: Department of Anatomy and Cell Biology, Université de Sherbrooke , 3201 Jean-Mignault, Sherbrooke, Québec J1E 4K8, Canada.
authors: Dominique
authors: D
authors: Lévesque
authors: Department of Anatomy and Cell Biology, Université de Sherbrooke , 3201 Jean-Mignault, Sherbrooke, Québec J1E 4K8, Canada.
authors: François-Michel
authors: FM
authors: Boisvert
mesh_terms: D018797:Cell Cycle Proteins / Q000378:metabolism; D002846:Chromatography, Affinity; D004249:DNA Damage; D004260:DNA Repair; D004261:DNA Replication; D006801:Humans; D013058:Mass Spectrometry; D064110:Minichromosome Maintenance Proteins / Q000378:metabolism / Q000502:physiology*; D011485:Protein Binding; D060066:Protein Interaction Maps / Q000502:physiology*
chemical_list: D018797:Cell Cycle Proteins; D064110:Minichromosome Maintenance Proteins
keywords: AP−MS; BioID; DNA damage; DNA replication; MCM complex; SILAC; mass spectrometry