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Status |
Public on Dec 31, 2020 |
Title |
Capture and identification of the eIF4E:mRNA interactome in human cells |
Organism |
Homo sapiens |
Experiment type |
Expression profiling by high throughput sequencing
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Summary |
Here we develop a novel methodology, capCLIP, to capture and identify mRNA interactions with the major cellular cap-binding protein eIF4E.
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Overall design |
HeLa cells in which a 3X-flag epitope tag has been introduced on to the N-terminal end of endogenous eIF4E protein were treated with either 100 nM rapamycin (or DMSO carrier along) for 2 hours, and then subject to 350 mJ/cm2 of UV irradiation to crosslink eIF4E to the m7G cap of mRNA. Biological replicates have been done for each treatment condition. Hela cells in which a 3X-flag epitope tag has been introduced on to the N-terminal end of endogenous eIF4E protein were serum-starved overnight then pre-treated for 1 hour with either 100 nM eFT-508 or with vehicle DMSO. Cells were then stimulated for 30 minutes with 500 nM PMA to activate MAP kinase signalling and thus MNK activity, and then subject to 350 mJ/cm2 of UV irradiation to crosslink eIF4E to the m7G cap of mRNA. Biological replicates have been done for each treatment condition.
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Contributor(s) |
Jensen K, Toubia J |
Citation(s) |
34255842 |
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Submission date |
Oct 06, 2019 |
Last update date |
Aug 09, 2021 |
Contact name |
Kirk B Jensen |
E-mail(s) |
kirk.jensen@sahmri.com
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Organization name |
South Australian Health & Medical Research Institute
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Department |
Lifelong Health
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Lab |
Cell Signalling & Gene Expression Group
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Street address |
North Terrace
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City |
Adelaide |
State/province |
South Australia |
ZIP/Postal code |
5000 |
Country |
Australia |
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Platforms (1) |
GPL18573 |
Illumina NextSeq 500 (Homo sapiens) |
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Samples (8)
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GSM4109521 |
flag-eIF4E HeLa cells; DMSO only; biological replicate #1 |
GSM4109522 |
flag-eIF4E HeLa cells; DMSO only; biological replicate #2 |
GSM4109523 |
flag-eIF4E HeLa cells; rapamycin treatment; biological replicate #1 |
GSM4109524 |
flag-eIF4E HeLa cells; rapamycin treatment; biological replicate #2 |
GSM5508128 |
flag-eIF4E Hela cells; serum starved overnight; PMA stimuation only; biological replicate #1 |
GSM5508129 |
flag-eIF4E Hela cells; serum starved overnight; PMA stimuation only; biological replicate #2 |
GSM5508130 |
flag-eIF4E Hela cells; serum starved overnight; eFT-508 pre-treat followed by PMA stimuation; biological replicate #1 |
GSM5508131 |
flag-eIF4E Hela cells; serum starved overnight; eFT-508 pre-treat followed by PMA stimuation; biological replicate #2 |
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Relations |
BioProject |
PRJNA576078 |
SRA |
SRP224550 |