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Diversified Application of barcoded PLATO(PLATO-BC) Platform for Identification of Protein Interactions

Genomics, Proteomics & Bioinformatics 2019年第3期17卷 319-331页

作者： Weili Kong Tsuyoshi Hayashi Guillaume Fiches Qikai Xu Mamie ZLi Jianwen Que Shuai Liu Wei Zhang Jun Qi Netty Santoso Stephen JElledge Jian Zhu Department of Microbiology and Immunology Department of Biochemistry and BiophysicsUniversity of Rochester Medical Center Division of Genetics Brigham and Women’s HospitalHoward Hughes Medical InstituteDepartment of GeneticsHarvard Medical School Department of Medicine Columbia University Medical Center Department of Chemistry College of Science and MathematicsUniversity of Massachusetts Boston Dana-Farber Cancer Institute Harvard Medical School Department of Pathology Ohio State University Wexner Medical Center

Proteins usually associate with other molecules physically to execute their *** these interactions is important for the functional analysis of ***,we reported the parallel analysis of translated ORFs(PLATO)to couple ribosome display of full-length ORFs with affinity enrichment of mRNA/protein/ribosome complexes for the “bait”molecules,followed by the deep sequencing analysis of ***,the sample processing,from extraction of precipitated mRNA to generation of DNA libraries,includes numerous steps,which is tedious and may cause the loss of *** PLATO(PLATO-BC),an improved platform was further developed to test its application for protein interaction *** this report,we tested the antisera-antigen interaction using serum samples from patients with inclusion body myositis(IBM).Tripartite motif containing 21(TRIM21)was identified as a potentially new IBM *** also expanded the application of PLATO-BC to identify protein interactions for JQ1,single ubiquitin peptide,and NS5 protein of Zika *** PLATO-BC analyses,we identified new protein interactions for these “bait”*** demonstrate that Ewing sarcoma breakpoint region 1(EWSR1)binds to JQ1 and their interactions may interrupt the EWSR1 association with acetylated histone *** kinase 3(RIOK3),a newly identified ubiquitin-binding protein,is preferentially associated with K63-ubiquitin *** also find that Zika NS5 protein interacts with two previously unreported host proteins,par-3 family cell polarity regulator(PARD3)and chromosome 19 open reading frame 53(C19orf53),whose attenuated expression benefits the replication of Zika *** results further demonstrate that PLATO-BC is capable of identifying novel protein interactions for various types of “bait”molecules.

关键词： barcoded PLATO Protein interaction Ubiquitin-binding protein Bromodomain inhibitor JQ1 Zika virus

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