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Status |
Public on Apr 30, 2015 |
Title |
Sensing the end: how TUT7 controls the fate of precursor microRNAs by uridylation |
Organism |
Homo sapiens |
Experiment type |
Non-coding RNA profiling by high throughput sequencing
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Summary |
Terminal uridylyl transferases (TUTs) function as integral regulators of microRNA (miRNA) biogenesis by modifying the end structure of precursor miRNA (pre-miRNA). Using biochemistry and deep sequencing techniques, we here investigate the mechanism how human TUT7 recognizes and uridylates pre-miRNAs. We show that the overhang of a pre-miRNA is the key structural element that TUT7 and its paralogues, TUT4 and TUT2, recognize. For group II pre-miRNAs which have a 1 nt 3’ overhang, TUT7 restores the canonical end structure (2 nt 3’ overhang) by mono-uridylation, and thereby promotes miRNA biogenesis. Interestingly, once the 3’ end is receded into the stem (3’ trimmed pre-miRNAs such as Ago-cleaved-pre-miRNA), TUT7 effectively generates an oligo-U tail that consequently leads to degradation. Our single-molecule study further suggests that a distributive mode is employed for both pathways, but the overhang length determines the frequency of TUT7-RNA interaction. Our results explain how TUT7 and TUT4 differentiate pre-miRNA species and reveal a role for TUT7 and TUT4 in the oligo-uridylation and removal of defective pre-miRNAs.
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Overall design |
HeLa cells were knocked down of control or TUT2/4/7, then total RNAs were prepared for RNA-seq
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Contributor(s) |
Kim B, Chang H, Kim VN |
Citation(s) |
25979828 |
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Submission date |
Dec 23, 2014 |
Last update date |
May 15, 2019 |
Contact name |
Hyeshik Chang |
E-mail(s) |
[email protected]
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Organization name |
Seoul National University
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Department |
School of Biological Sciences
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Lab |
Hyeshik Chang Lab
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Street address |
Building 203 Room 525, School of Biological Sciences, Seoul National University, 1 Gwanak-ro, Gwanak-gu
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City |
Seoul |
State/province |
South Korea |
ZIP/Postal code |
08826 |
Country |
South Korea |
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Platforms (1) |
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Samples (2) |
GSM1572155 |
HeLa transfected with a control siRNA |
GSM1572156 |
HeLa transfected with siRNAs for TUT2, 4, 7 |
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Relations |
BioProject |
PRJNA271095 |
SRA |
SRP051515 |
Supplementary file |
Size |
Download |
File type/resource |
GSE64482_RAW.tar |
1.9 Mb |
(http)(custom) |
TAR (of TXT) |
SRA Run Selector |
Raw data are available in SRA |
Processed data provided as supplementary file |
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