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| Status |
Public on May 04, 2021 |
| Title |
NUDT21 regulates intron detention of the SAM synthetase MAT2A RNA |
| Organism |
Homo sapiens |
| Experiment type |
Expression profiling by high throughput sequencing
|
| Summary |
NUDT21 encodes the CFIm25 protein, a component of the CFIm complex that regulates alternative polyadenylation (APA). Our data suggest that the CFIm complex also induces splicing of a detained intron in the the S-adenosylmethionine (SAM) synthetase MAT2A to control intracellular levels of SAM. To genetically separate these functions, we used poly(A) Click-seq (PAC-seq) to determine the changes in poly(A) site selection in two cell lines. The parental cell line is the colorectal cell line HCT116 and a derivative line that has the detained intron of MAT2A deleted in both alleles of MAT2A (116-DDI). We found few differences in alternative polyadenylation after CFIm25 depletion were similar supporting teh conclusion that CFIm's role in APA is mechanistically distinct from its role in regulation of MATA splicing.
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| Overall design |
We used siRNAs to knockdown NUDT21/CFIm25 expression for four days in either the parental HCT116 cell line or in 116-ΔDI cells. A nontargeting (NT) siRNA was used as a control. We then performed PAC-seq with ClickSeq Technologies (Galveston, TX) to analyze changes in APA. Three independent biological replicates were performed.
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| Contributor(s) |
Conrad NK, Scarborough AM |
| Citation(s) |
33949310 |
| NIH grant(s) |
| Grant ID |
Grant title |
Affiliation |
Name |
| R01 GM127311 |
Mechanisms of posttranscriptional regulation of SAM homeostasis |
UT SOUTHWESTERN MEDICAL CENTER |
NICHOLAS K CONRAD |
|
| Submission date |
Sep 25, 2020 |
| Last update date |
Aug 03, 2021 |
| Contact name |
Nicholas K Conrad |
| E-mail(s) |
nicholas.conrad@utsouthwestern.edu
|
| Phone |
2146480795
|
| Organization name |
UT Southwestern Medical Center
|
| Department |
Microbiology
|
| Street address |
6000 Harry Hines Blvd
|
| City |
Dallas |
| State/province |
TX |
| ZIP/Postal code |
75063 |
| Country |
USA |
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| Platforms (1) |
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| Samples (12)
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| Relations |
| BioProject |
PRJNA665731 |
| SRA |
SRP285411 |
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