title: Random mutagenesis MAPPIT analysis identifies binding sites for Vif and Gag in both cytidine deaminase domains of Apobec3G.
abstract: 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.
authors.affiliation: Department of Medical Protein Research, VIB, Ghent, Belgium.
authors.forename: Isabel
authors.initials: I
authors.lastname: Uyttendaele
authors.forename: Delphine
authors.initials: D
authors.lastname: Lavens
authors.forename: Dominiek
authors.initials: D
authors.lastname: Catteeuw
authors.forename: Irma
authors.initials: I
authors.lastname: Lemmens
authors.forename: Celia
authors.initials: C
authors.lastname: Bovijn
authors.forename: Jan
authors.initials: J
authors.lastname: Tavernier
authors.forename: Frank
authors.initials: F
authors.lastname: Peelman
mesh_terms: D000071480:APOBEC-3G Deaminase; D001665:Binding Sites; D003564:Cytidine Deaminase / Q000737:chemistry* / Q000378:metabolism; D057809:HEK293 Cells; D015497:HIV-1 / Q000378:metabolism; D006801:Humans; D008958:Models, Molecular; D016296:Mutagenesis; D050505:Mutant Proteins / Q000378:metabolism; D009154:Mutation / Q000235:genetics; D011485:Protein Binding; D025941:Protein Interaction Mapping; D055503:Protein Multimerization; D017434:Protein Structure, Tertiary; D020798:Two-Hybrid System Techniques; D054301:gag Gene Products, Human Immunodeficiency Virus / Q000378:metabolism*; D054320:vif Gene Products, Human Immunodeficiency Virus / Q000378:metabolism*
chemical_list: D050505:Mutant Proteins; D054301:gag Gene Products, Human Immunodeficiency Virus; D054320:vif Gene Products, Human Immunodeficiency Virus; D000071480:APOBEC-3G Deaminase; C464924:APOBEC3G protein, human; D003564:Cytidine Deaminase