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| Status |
Public on Mar 01, 2022 |
| Title |
Oppositional Poly(A) Tail Length Regulation by FMRP and CPEB1 |
| Organism |
Mus musculus |
| Experiment type |
Expression profiling by high throughput sequencing
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| Summary |
Poly(A) tail length is regulated in both the nucleus and cytoplasm. One factor that controls polyadenylation in the cytoplasm is CPEB1, an RNA binding protein that associates with specific mRNA 3’UTR sequences to tether enzymes that add and remove poly(A). Two of these enzymes, the noncanonical poly(A) polymerases Gld2 (TENT2, PAPD4, Wispy) and Gld4 (TENT4B, PAPD5, TRF4, TUT3), interact with CPEB1 to extend poly(A). To identify additional RNA binding proteins that might anchor Gld4 to RNA, we expressed double tagged Gld4 in U87MG cells, which was used for sequential immunoprecipitation and elution followed by mass spectrometry. We identified several RNA binding proteins that co-precipitated with Gld4, among which was FMRP. To assess whether FMRP regulates polyadenylation, we performed TAIL-seq from WT and FMRP-deficient HEK293 cells. Surprisingly, loss of FMRP resulted in an overall increase in poly(A), which was also observed for several specific mRNAs. Conversely, loss of CPEB1 elicited an expected decrease in poly(A), which was examined in cultured neurons. We also examined polyadenylation in wild type (WT) and FMRP-deficient mouse brain cortex by direct RNA Nanopore™ sequencing, which identified RNAs with both increased and decreased poly(A). Our data show that FMRP has a role in mediating poly(A) tail length, which adds to its repertoire of RNA regulation.
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| Overall design |
Identification of Poly(A) tail length for all transcripts in WT and FMRP Knock-out mice using direct RNA sequencing (PromethION)
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| Contributor(s) |
Shin J, Paek KY, Chang H, Chikhaoui L, Jung S, Pony S, Suzuki Y, Kim VN, Padmanabhan K, Richter JD |
| Citation(s) |
35217597 |
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| Submission date |
Nov 15, 2021 |
| Last update date |
Mar 01, 2022 |
| Contact name |
Kiran Padmanabhan |
| E-mail(s) |
kiran.padmanabhan@ens-lyon.fr
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| Organization name |
ENS-Lyon
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| Department |
IGFL
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| Street address |
32-34 avenue Tony Garnier
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| City |
Lyon |
| ZIP/Postal code |
69007 |
| Country |
France |
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| Platforms (1) |
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| Samples (6)
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| Relations |
| BioProject |
PRJNA780496 |
| SRA |
SRP346148 |
| Supplementary file |
Size |
Download |
File type/resource |
| GSE188840_Mouse_Brain_Cortex_Results.xlsx |
24.7 Mb |
(ftp)(http) |
XLSX |
SRA Run Selector |
| Raw data are available in SRA |
| Processed data are available on Series record |
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