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Multiple competing RNA structures dynamically control alternative splicing in the human ATE1 gene.

Authors
  • Kalinina, Marina1
  • Skvortsov, Dmitry2
  • Kalmykova, Svetlana1
  • Ivanov, Timofei1
  • Dontsova, Olga1, 2
  • Pervouchine, Dmitri D1
  • 1 Skolkovo Institute of Science and Technology, Center of Life Sciences, Moscow 143026, Russia.
  • 2 Moscow State University, Faculty of Chemistry, Moscow 119991, Russia.
Type
Published Article
Journal
Nucleic Acids Research
Publisher
Oxford University Press
Publication Date
Jan 11, 2021
Volume
49
Issue
1
Pages
479–490
Identifiers
DOI: 10.1093/nar/gkaa1208
PMID: 33330934
Source
Medline
Language
English
License
Unknown

Abstract

The mammalian Ate1 gene encodes an arginyl transferase enzyme with tumor suppressor function that depends on the inclusion of one of the two mutually exclusive exons (MXE), exons 7a and 7b. We report that the molecular mechanism underlying MXE splicing in Ate1 involves five conserved regulatory intronic elements R1-R5, of which R1 and R4 compete for base pairing with R3, while R2 and R5 form an ultra-long-range RNA structure spanning 30 Kb. In minigenes, single and double mutations that disrupt base pairings in R1R3 and R3R4 lead to the loss of MXE splicing, while compensatory triple mutations that restore RNA structure revert splicing to that of the wild type. In the endogenous Ate1 pre-mRNA, blocking the competing base pairings by LNA/DNA mixmers complementary to R3 leads to the loss of MXE splicing, while the disruption of R2R5 interaction changes the ratio of MXE. That is, Ate1 splicing is controlled by two independent, dynamically interacting, and functionally distinct RNA structure modules. Exon 7a becomes more included in response to RNA Pol II slowdown, however it fails to do so when the ultra-long-range R2R5 interaction is disrupted, indicating that exon 7a/7b ratio depends on co-transcriptional RNA folding. In sum, these results demonstrate that splicing is coordinated both in time and in space over very long distances, and that the interaction of these components is mediated by RNA structure. © The Author(s) 2020. Published by Oxford University Press on behalf of Nucleic Acids Research.

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