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Series GSE132009 Query DataSets for GSE132009
Status Public on Dec 03, 2019
Title Suppression of ribosomal pausing by eIF5A is necessary to maintain the fidelity of start codon selection (CRISPR screen)
Organism Homo sapiens
Experiment type Other
Summary Sequences within 5' untranslated regions (UTRs) dictate the site and efficiency of translation initiation. In this study, an unbiased screen designed to interrogate the 5' UTR-mediated regulation of the growth-promoting gene MYC unexpectedly revealed the ribosomal pause-relief factor eIF5A as a regulator of translation initiation codon selection. Depletion of eIF5A enhanced upstream translation within 5' UTRs across yeast and human transcriptomes, including on the MYC transcript where this resulted in increased production of an N-terminally extended protein. Furthermore, ribosome profiling experiments established that the function of eIF5A as a suppressor of ribosomal pausing at sites of suboptimal peptide bond formation is conserved in human cells. We present evidence that proximal ribosomal pausing on a transcript triggers enhanced usage of upstream suboptimal or non-canonical initiation codons. Thus, we propose that eIF5A functions not only to maintain efficient translation elongation in eukaryotic cells, but also to maintain the fidelity of translation initiation.
 
Overall design A genome-scale CRISPR/Cas9-mediated loss of function screen was performed in HCT116 cells expressing either a MYC 5' UTR EGFP reporter or a control EGFP reporter.
 
Contributor(s) Manjunath H, Zhang H, Mendell J
Citation(s) 31801078
Submission date May 31, 2019
Last update date Dec 31, 2019
Contact name Joshua Mendell
E-mail(s) joshua.mendell@utsouthwestern.edu
Organization name UT Southwestern Medical Center
Department Molecular Biology
Street address 6000 Harry Hines Blvd., NA6.200A
City Dallas
State/province TX
ZIP/Postal code 75390-9148
Country USA
 
Platforms (1)
GPL18573 Illumina NextSeq 500 (Homo sapiens)
Samples (16)
GSM3834408 HCT116_EGFP_Lib A_Unsorted_Replicate 1 (HM9)
GSM3834409 HCT116_EGFP_Lib B_Unsorted_Replicate 1 (HM10)
GSM3834410 HCT116_EGFP_Lib A_Bright_Replicate 1 (HM1)
This SubSeries is part of SuperSeries:
GSE132010 Suppression of ribosomal pausing by eIF5A is necessary to maintain the fidelity of start codon selection
Relations
BioProject PRJNA545621
SRA SRP199982

Download family Format
SOFT formatted family file(s) SOFTHelp
MINiML formatted family file(s) MINiMLHelp
Series Matrix File(s) TXTHelp

Supplementary file Size Download File type/resource
GSE132009_MYC-UTR.norm.count_updated_.txt.gz 3.6 Mb (ftp)(http) TXT
SRA Run SelectorHelp
Raw data are available in SRA
Processed data are available on Series record

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