title: Screening of binding proteins that interact with human Salvador 1 in a human fetal liver cDNA library by the yeast two-hybrid system.
abstract: Salvador promotes both cell cycle exit and apoptosis through the modulation of both cyclin E and Drosophila inhibitor of apoptosis protein in Drosophila. However, the cellular function of human Salvador (hSav1) is rarely reported. To screen for novel binding proteins that interact with hSav1, the cDNA of hSav1 was cloned into a bait protein plasmid, and positive clones were screened from a human fetal liver cDNA library by the yeast two-hybrid system. hSav1 mRNA was expressed in yeast and there was no self-activation and toxicity in the yeast strain AH109. Twenty proteins were found to interact with hSav1, including HS1 (haematopoietic cell specific protein1)-associated protein X-1 (HAX-1); neural precursor cell expressed, developmentally down-regulated 9, pyruvate kinase, liver and RBC, cytochrome c oxidase subunit Vb, enoyl coenzyme A hydratase short chain 1, and NADH dehydrogenase (ubiquinone) 1 beta subcomplex, demonstrating that the yeast two-hybrid system is an efficient method for investigating protein interactions. Among the identified proteins, there were many mitochondrial proteins, indicating that hSav1 may play a role in mitochondrial function. We also confirmed the interaction of HAX-1 and hSav1 in mammalian cells. This investigation provides functional clues for further exploration of potential apoptosis-related proteins in disease biotherapy.
authors.affiliation: Institute of Molecular Medicine Center, Tongji Hospital, Tongji Medical College, Huazhong University of Science and Technology, Wuhan 430030, People's Republic of China.
authors.forename: Xiaolan
authors.initials: X
authors.lastname: Li
authors.forename: Xuelai
authors.initials: X
authors.lastname: Luo
authors.forename: Zhaoming
authors.initials: Z
authors.lastname: Li
authors.forename: Guihua
authors.initials: G
authors.lastname: Wang
authors.forename: Hui
authors.initials: H
authors.lastname: Xiao
authors.forename: Deding
authors.initials: D
authors.lastname: Tao
authors.forename: Jianping
authors.initials: J
authors.lastname: Gong
authors.forename: Junbo
authors.initials: J
authors.lastname: Hu
mesh_terms: D048868:Adaptor Proteins, Signal Transducing / Q000378:metabolism*; D002352:Carrier Proteins / Q000235:genetics / Q000378:metabolism*; D018797:Cell Cycle Proteins / Q000235:genetics / Q000378:metabolism*; D002460:Cell Line; D015723:Gene Library; D006801:Humans; D010957:Plasmids; D011485:Protein Binding; D025941:Protein Interaction Mapping; D017422:Sequence Analysis, DNA; D020798:Two-Hybrid System Techniques
chemical_list: D048868:Adaptor Proteins, Signal Transducing; D002352:Carrier Proteins; D018797:Cell Cycle Proteins; C104759:HAX1 protein, human; C415864:SAV1 protein, human