|
| Status |
Public on Nov 07, 2022 |
| Title |
DAP5 enables main ORF translation on mRNAs with structured and uORF-containing 5’ leaders |
| Organism |
Homo sapiens |
| Experiment type |
Expression profiling by high throughput sequencing
Other
|
| Summary |
Half of mammalian transcripts contain short upstream open reading frames (uORFs) that potentially regulate translation of the downstream coding sequence (CDS). The molecular mechanisms governing these events remain poorly understood. Here, we find that the non-canonical initiation factor Death-associated protein 5 (DAP5 or eIF4G2) is required for translation initiation on select transcripts. Using ribosome profiling and luciferase-based reporters coupled with mutational analysis we show that DAP5-dependent translation occurs on messenger RNAs (mRNAs) with long, structure-prone 5′ leader sequences and persistent uORF translation. These mRNAs preferentially code for signalling factors such as kinases and phosphatases. We also report that cap/eIF4F- and eIF4A-dependent recruitment of DAP5 to the mRNA facilitates main CDS, but not uORF, translation suggesting a role for DAP5 in translation re-initiation. Our study reveals important mechanistic insights into how a non-canonical translation initiation factor involved in stem cell fate shapes the synthesis of specific signalling factors.
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| |
|
| Overall design |
Two cell types with two biological replicates analysed using RNA-Seq and Ribo-Seq
|
| |
|
| Contributor(s) |
Weber R, Igreja C |
| Citation(s) |
36473845 |
| |
| Submission date |
Aug 07, 2020 |
| Last update date |
Jan 04, 2023 |
| Contact name |
Ramona Weber |
| E-mail(s) |
ramona.weber@uzh.ch
|
| Organization name |
University of Zurich
|
| Department |
Institute for Regenerative Medicine
|
| Street address |
Wagistrasse 12
|
| City |
Schlieren |
| ZIP/Postal code |
8952 |
| Country |
Switzerland |
| |
|
| Platforms (1) |
| GPL21290 |
Illumina HiSeq 3000 (Homo sapiens) |
|
| Samples (8)
|
| GSM4712999 |
control-1_RNA: Wild-type replicate 1 for RNA-seq |
| GSM4713000 |
control-2_RNA: Wild-type replicate 2 for RNA-seq |
| GSM4713001 |
DAP5-1_RNA: DAP5-null replicate 1 for RNA-seq |
| GSM4713002 |
DAP5-2_RNA: DAP5-null replicate 2 for RNA-seq |
| GSM4713003 |
control-1_Ribo: Wild-type replicate 1 for Ribo-seq |
| GSM4713004 |
control-2_Ribo: Wild-type replicate 2 for Ribo-seq |
| GSM4713005 |
DAP5-1_Ribo: DAP5-null replicate 1 for Ribo-seq |
| GSM4713006 |
DAP5-2_Ribo: DAP5-null replicate 2 for Ribo-seq |
|
| Relations |
| BioProject |
PRJNA655835 |
| SRA |
SRP276619 |
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