Random mutagenesis MAPPIT analysis identifies binding sites for Vif and Gag in both cytidine deaminase domains of Apobec3G

• Published in: PloS one 2012-09, Vol.7 (9), p.e44143
• Main Authors: Uyttendaele, Isabel, Lavens, Delphine, Catteeuw, Dominiek, Lemmens, Irma, Bovijn, Celia, Tavernier, Jan, Peelman, Frank
• Format: Article
• Language: English
• Subjects: Amino acids; Analysis; APOBEC-3G Deaminase; Binding Sites; Biochemistry; Biology; Cells (Biology); Crystallography; Cytidine deaminase; Cytidine Deaminase - chemistry; Cytidine Deaminase - metabolism; Cytokines; Dimerization; Enzymes; gag Gene Products, Human Immunodeficiency Virus - metabolism; Health sciences; HEK293 Cells; HIV; HIV-1 - metabolism; Human immunodeficiency virus; Humans; Hybrid systems; Medicine; Methods; Microscopy; Models, Molecular; Mutagenesis; Mutant Proteins - metabolism; Mutation; Mutation - genetics; Protein Binding; Protein interaction; Protein Interaction Mapping; Protein Multimerization; Protein Structure, Tertiary; Protein-protein interactions; Proteins; Random mutagenesis; Strategy; Two-Hybrid System Techniques; vif Gene Products, Human Immunodeficiency Virus - metabolism
• ISSN: 1932-6203; 1932-6203
• DOI: 10.1371/journal.pone.0044143

The mammalian two-hybrid system MAPPIT allows the detection of protein-protein interactions in intact human cells. We developed a random mutagenesis screening strategy based on MAPPIT to detect mutations that disrupt the interaction of one protein with multiple protein interactors simultaneously. The strategy was used to detect residues of the human cytidine deaminase Apobec3G that are important for its homodimerization and its interaction with the HIV-1 Gag and Vif proteins. The strategy is able to identify the previously described head-to-head homodimerization interface in the N-terminal domain of Apobec3G. Our analysis further detects two new potential interaction surfaces in the N-and C-terminal domain of Apobec3G for interaction with Vif and Gag or for Apobec3G dimerization.