title: Identification of PLP2 and RAB5C as novel TPD52 binding partners through yeast two-hybrid screening.
abstract: Tumor protein D52 (TPD52) is overexpressed in different cancers, but its molecular functions are poorly defined. A large, low-stringency yeast two-hybrid screen using full-length TPD52 bait identified known partners (TPD52, TPD52L1, TPD52L2, MAL2) and four other preys that reproducibly bound TPD52 and TPD52L1 baits (PLP2, RAB5C, GOLGA5, YIF1A). PLP2 and RAB5 interactions with TPD52 were confirmed in pull down assays, with interaction domain mapping experiments indicating that both proteins interact with a novel binding region of TPD52. This study provides insights into TPD52 functions, and ways to maximise the efficiency of low-stringency yeast two-hybrid screens.
authors.affiliation: Children's Cancer Research Unit, Kids Research Institute, The Children's Hospital at Westmead, Locked Bag 4001, Westmead, NSW, 2145, Australia.
authors.forename: Hamideh
authors.initials: H
authors.lastname: Shahheydari
authors.forename: Sarah
authors.initials: S
authors.lastname: Frost
authors.forename: Brian J
authors.initials: BJ
authors.lastname: Smith
authors.forename: Guy E
authors.initials: GE
authors.lastname: Groblewski
authors.forename: Yuyan
authors.initials: Y
authors.lastname: Chen
authors.forename: Jennifer A
authors.initials: JA
authors.lastname: Byrne
mesh_terms: D001665:Binding Sites; D045744:Cell Line, Tumor; D004926:Escherichia coli / Q000235:genetics / Q000378:metabolism; D015870:Gene Expression; D006801:Humans; D062792:MARVEL Domain-Containing Proteins / Q000737:chemistry / Q000235:genetics / Q000378:metabolism*; D009363:Neoplasm Proteins / Q000737:chemistry / Q000235:genetics / Q000378:metabolism*; D010957:Plasmids / Q000378:metabolism; D011485:Protein Binding; D054730:Protein Interaction Domains and Motifs; D025941:Protein Interaction Mapping; D011510:Proteolipids / Q000737:chemistry / Q000235:genetics / Q000378:metabolism*; D011993:Recombinant Fusion Proteins / Q000737:chemistry / Q000235:genetics / Q000378:metabolism*; D012441:Saccharomyces cerevisiae / Q000235:genetics / Q000378:metabolism; D014162:Transfection; D020798:Two-Hybrid System Techniques; D020696:rab5 GTP-Binding Proteins / Q000737:chemistry / Q000235:genetics / Q000378:metabolism*
chemical_list: D062792:MARVEL Domain-Containing Proteins; D009363:Neoplasm Proteins; C094765:PLP2 protein, human; D011510:Proteolipids; D011993:Recombinant Fusion Proteins; C114751:TPD52 protein, human; C427755:RAB5C protein, human; D020696:rab5 GTP-Binding Proteins