Title
Diversified Application of Barcoded PLATO (PLATO-BC) Platform for Identification of Protein Interactions.
Abstract
Proteins usually associate with other molecules physically to execute their functions. Identifying these interactions is important for the functional analysis of proteins. Previously, 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 mRNA. However, 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 materials. Barcoded PLATO (PLATO-BC), an improved platform was further developed to test its application for protein interaction discovery. In 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 autoantigen. We also expanded the application of PLATO-BC to identify protein interactions for JQ1, single ubiquitin peptide, and NS5 protein of Zika virus. From PLATO-BC analyses, we identified new protein interactions for these “bait” molecules. We demonstrate that Ewing sarcoma breakpoint region 1 (EWSR1) binds to JQ1 and their interactions may interrupt the EWSR1 association with acetylated histone H4. RIO kinase 3 (RIOK3), a newly identified ubiquitin-binding protein, is preferentially associated with K63-ubiquitin chain. We 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 virus. These results further demonstrate that PLATO-BC is capable of identifying novel protein interactions for various types of “bait” molecules.
Year
DOI
Venue
2019
10.1016/j.gpb.2018.12.010
Genomics, Proteomics & Bioinformatics
Keywords
Field
DocType
Barcoded PLATO,Protein interaction,Ubiquitin-binding protein,Bromodomain inhibitor JQ1,Zika virus
Ribosome display,Protein–protein interaction,Biology,Open reading frame,Messenger RNA,Ribosome,EWING SARCOMA BREAKPOINT REGION 1,ORFS,Genetics,Histone H4
Journal
Volume
Issue
ISSN
17
3
1672-0229
Citations 
PageRank 
References 
0
0.34
0
Authors
12
Name
Order
Citations
PageRank
Weili Kong100.34
Tsuyoshi Hayashi200.34
Guillaume Fiches300.34
Qikai Xu400.34
Mamie Li500.34
Jianwen Que600.34
Shuai Liu700.34
Wei Zhang800.34
Jun Qi900.34
Netty Santoso1000.34
Stephen J Elledge1100.34
Jian Zhu1200.34