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Comprehensive Identification of FXR1 Targets Using pSILAC-BONCAT Proteomics

使用 pSILAC-BONCAT 蛋白质组学全面鉴定 FXR1 靶标

基本信息

批准号：
9241667
负责人：
Viswanathan Palanisamy
金额：
$ 22.43万
依托单位：
MEDICAL UNIVERSITY OF SOUTH CAROLINA
依托单位国家：
美国
项目类别：
财政年份：
2017
资助国家：
美国
起止时间：
2017-01-01 至 2018-12-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/9241667
关键词：
Affect Amino Acids Apoptosis Architecture Bioinformatics Biological Biological Assay Biological Process Bypass CDKN1A gene Cell Aging Cell Culture Techniques Cell Cycle Arrest Cell Line Cells Chemistry Clinic Data Development Disease Dissection Embryo Eukaryotic Cell Event FMR1 FXR1 gene Family Fibroblasts Foundations Fragile X Syndrome G1 Phase Gene Expression Gene Expression Alteration Gene Expression Profile Genes Genetic Genetic Transcription Genetic Translation Goals Head and Neck Cancer Human Immunoprecipitation Laboratories Malignant - descriptor Malignant Epithelial Cell Malignant Neoplasms Mass Spectrum Analysis Mediating Messenger RNA Molecular Molecular Profiling Mouth Carcinoma Mus Non-Small-Cell Lung Carcinoma Ontology Oral Pathway interactions Pattern Phenotype Physiologic pulse Play Polyribosomes Post-Transcriptional Regulation Predisposition Prevalence Proteins Proteome Proteomics Publishing RNA RNA Processing RNA-Binding Proteins Recruitment Activity Regulation Regulator Genes Reporting Research Role Stable Isotope Labeling Stimulus Stress Survival Rate Systems Biology TP53 gene Telomerase RNA Component Testing Transcription Process Translating Translations Tumor Tissue Validation Western Blotting aggressive therapy base cancer cell carcinogenesis conventional therapy experience genetic signature genome-wide head and neck cancer patient high throughput analysis keratinocyte knock-down loss of function mRNA Decay mRNA Stability malignant mouth neoplasm mouth squamous cell carcinoma mutant neoplastic cell new therapeutic target novel marker overexpression prevent protein expression screening senescence small hairpin RNA targeted treatment therapeutic target therapy resistant tumor tumor heterogeneity tumorigenesis

项目摘要

The purpose of this project is to determine the post-transcriptional regulation that changes the progression of oral cancer via the RNA-binding protein FXR1 and its mRNA translational control that govern cellular senescence. FXR1 was considered as a critical regulator of gene expression in most of the eukaryotic cells by controlling mRNA stability and translation. Thus, the main goal of this project is to understand the molecular mechanisms of FXR1-mediated mRNA translation and their implications in oral squamous cell carcinoma (OSCC) cellular senescence. Cellular senescence is an important mechanism for preventing the proliferation of potential cancer cells. Hence, underpinning the mechanistic aspects of cellular machinery by dissection the protein expression profiles provides novel biomarkers and/or therapeutic targets. Our laboratory has extensive experience in studying RNA-binding proteins and high throughput analysis of protein expression by pulsed-stable isotope labeling with amino acids in cell culture (pSILAC). We have developed a genome- wide shRNA screen of RBPs to test the cellular senescence in oral cancer cells. By studying oral cancer cells for cellular senescence, we have discovered FXR1 is overexpressed and depletion of FXR1 induces cellular senescence. Interestingly, FXR1 is a major player to control the expression of protein at the translational level that could mediate cellular senescence. Hence, we plan to systematically study the phenotypic changes of cellular senescence in oral cancer cells by understanding the expression of proteins that are controlled by FXR1. Thus, our central hypothesis is to understand how overexpression of FXR1 in oral cancer cells alter the protein expression by modulating RNA processing machinery to control cellular senescence. We propose the following specific aims to test this hypothesis: 1) to determine the critical role of FXR1 in bypassing OSCC senescence. We will test if the gain- and loss-of-function of FXR1 affects senescence in OSCC cells through various cell biological senescence assays, 2) to determine the consequence of FXR1 dysregulation on the cellular proteome. The protein targets of FXR1 will be identified using pulsed-Stable Isotope Labeling of Cells in cultured OSCC cells with click-chemistry Bioorthogonal Noncanonical Amino Acid Tagging (pSILAC- BONCAT) and Mass Spectrometry.

这个项目的目的是确定改变基因的转录后调控