Translation of small open reading frames in 3'UTR enhances translation of canonical open reading frames

3UTR 中小型开放阅读框的翻译增强了规范开放阅读框的翻译

• 批准号: 10671093
• 负责人: Qiushuang Wu
• 金额: $ 9.5万
• 依托单位: ROCKEFELLER UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2022
• 资助国家: 美国
• 起止时间: 2022-08-01 至 2026-07-31
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10671093
• 关键词: 3' Untranslated Regions; 5' Untranslated Regions; Affect; Award; Bioinformatics; Biological; Biological Assay; Cancer Biology; Cancer Etiology; Candidate Disease Gene; Cells; Cellular biology; Clinical; DNA; Data; Databases; Development; Disease; Drug Design; Drug Targeting; Embryo; Eukaryotic Cell; Future; Gene Expression; Gene Expression Regulation; Genes; Genetic Transcription; Genomics; Health; Human; Human Cell Line; Kidney Neoplasms; Kinetics; Knowledge; Light; Malignant Neoplasms; Measures; Mediating; Messenger RNA; Methodology; MicroRNAs; Molecular; Molecular Biology; Mutation; Open Reading Frames; Orthologous Gene; Outcome; Pathogenesis; Pathologic; Patients; Pattern; Peptide Initiation Factors; Peptides; Play; Polyadenylation; Population; Post-Transcriptional Regulation; Process; Proteins; Proteomics; Publishing; Regulation; Regulatory Element; Regulatory Pathway; Reporter; Reporting; Research; Research Project Grants; Ribosomes; Role; Site; Techniques; The Cancer Genome Atlas; Training; Translating; Translations; Untranslated RNA; Untranslated Regions; Vertebrates; Work; Zebrafish; base; cancer cell; career; genome-wide; genomic profiles; human disease; innovation; insight; interest; mRNA Stability; machine translation; molecular imaging; novel; novel diagnostics; ribosome profiling; single molecule; transcriptome sequencing; tumor

Project Summary Ribosome and proteomic profiling have revealed a large number of small translated open reading frames (ORF) within previously described “untranslated regions” (UTRs) and long non-coding RNAs. While some of the small ORFs depend on the encoded peptide to function in various fundamental processes (e.g., development). Translation of small ORFs in the 5’UTR, known as upstream-ORFs (uORFs), usually represses gene expression, independent of the encoded peptide. Small ORFs have also been reported in 3'UTR, termed downstream ORF (dORF). However, the dORF function and their relationship to human health and disease remain unknown. I characterized dORFs from human and zebrafish using ribosome profiling data. My preliminary data indicates, contrary to uORFs, translation of dORFs (small ORF in the 3’UTR) strongly enhances translation of the canonical ORFs and remains an uncharacterized regulatory mechanism across vertebrates. The objectives are: 1) Dissect at the single molecular level how dORF enhances translation of the canonical CDS. And 2) Determine whether alternative polyadenylation in cancer influences dORF regulation to cause cancer. The rationale for the proposed research is to gain a mechanistic understanding of dORF-mediated regulation and to assess the possible biological importance of dORF dysregulation under disease conditions (e.g. Cancer). This proposal is conceptually innovative as it is based on the exploration of a novel, yet widespread translation regulatory mechanism conserved across vertebrates. Technically, this proposal will combine single molecular imaging, genomic profiles (RNA-seq, Ribosome profiling), and reporter approaches in different human cell lines (including cancer cells) and published patient data. The outcomes from this project will emphasize the role of ribosome as a master gene expression regulator, and shield light on the importance of small ORFs. This translation kinetics work about dORF will provide critical insights into the molecular mechanism of this uncharacterized regulatory pathway. Exploring dORF dysregulation in cancer due to APA will highlight the mRNA itself as disease driver even without any mutation in DNA, and it also indicates possible clinical impact of dORF to detect and even cure cancer. My long-term interest is to study gene expression dysregulation in cancer. This training award will increase my knowledge background of cancer biology, molecular biology and bioinformatics. It will also promote the technical training of single molecular imaging, ribosome profiling, cell biology assays for cancer. Overall, this proposal will help me for future independent cancer molecular/genomic career.

项目总结