TENT2, TUT4, and TUT7 selectively regulate miRNA sequence and abundance

Acong Yang, Xavier Bofill- De Ros, Ryan Stanton, Tie-Juan Shao, Patricia Villanueva and Shuo Gu ()
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Acong Yang: National Cancer Institute
Xavier Bofill- De Ros: National Cancer Institute
Ryan Stanton: National Cancer Institute
Tie-Juan Shao: National Cancer Institute
Patricia Villanueva: National Cancer Institute
Shuo Gu: National Cancer Institute

Nature Communications, 2022, vol. 13, issue 1, 1-15

Abstract: Abstract TENTs generate miRNA isoforms by 3’ tailing. However, little is known about how tailing regulates miRNA function. Here, we generate isogenic HEK293T cell lines in which TENT2, TUT4 and TUT7 are knocked out individually or in combination. Together with rescue experiments, we characterize TENT-specific effects by deep sequencing, Northern blot and in vitro assays. We find that 3’ tailing is not random but highly specific. In addition to its known adenylation, TENT2 contributes to guanylation and uridylation on mature miRNAs. TUT4 uridylates most miRNAs whereas TUT7 is dispensable. Removing adenylation has a marginal impact on miRNA levels. By contrast, abolishing uridylation leads to dysregulation of a set of miRNAs. Besides let-7, miR-181b and miR-222 are negatively regulated by TUT4/7 via distinct mechanisms while the miR-888 cluster is upregulated specifically by TUT7. Our results uncover the selective actions of TENTs in generating 3’ isomiRs and pave the way to investigate their functions.

Date: 2022
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DOI: 10.1038/s41467-022-32969-8

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