PUBLICATION

Deep intron elements mediate nested splicing events at consecutive AG-dinucleotides to regulate alternative 3' splice site choice in vertebrate 4.1 genes

Authors
Parra, M.K., Gallagher, T.L., Amacher, S.L., Mohandas, N., and Conboy, J.G.
ID
ZDB-PUB-120409-5
Date
2012
Source
Molecular and cellular biology   32(11): 2044-2053 (Journal)
Registered Authors
Amacher, Sharon
Keywords
none
MeSH Terms
  • Alternative Splicing*
  • Animals
  • Humans
  • Introns*
  • Mice
  • Microfilament Proteins/genetics*
  • Microfilament Proteins/metabolism
  • RNA Precursors/genetics
  • RNA Precursors/metabolism
  • RNA Splice Sites*
  • Regulatory Sequences, Nucleic Acid*
PubMed
22473990 Full text @ Mol. Cell. Biol.
Abstract

Distal intraexon (iE) regulatory elements in 4.1R pre-mRNA govern 32 splice site choice at exon 2 (E2) via nested splicing events, ultimately modulating expression of N-terminal isoforms of cytoskeletal 4.1R protein. Here we explore intrasplicing in other normal and disease gene context(s) and demonstrate conservation of intrasplicing through vertebrate evolution. In the paralogous 4.1B gene, we identified <120kb upstream of E2 an ultra-distal intraexon, iEB, that mediates intrasplicing by promoting two intricately-coupled splicing events that ensure selection of a weak distal acceptor at E2 (E2dis) by prior excision of the competing proximal acceptor (E2prox). Mutating iEB in minigene splicing reporters abrogated intrasplicing, as did blocking endogenous iEB function with antisense morpholinos in live mouse and zebrafish animal models. In a human elliptocytosis patient with a mutant 4.1R gene lacking E2-E4, we showed that aberrant splicing is consistent with iER-mediated intrasplicing at the first available exons downstream of iER, namely alternative E5 and constitutive E6. Finally, analysis of heterologous acceptor contexts revealed a strong preference for nested 32 splice events at consecutive pairs of AG dinucleotides. Distal regulatory elements may control intrasplicing at a subset of alternative 32 splice sites in vertebrate pre-mRNAs to generate proteins with functional diversity.

Genes / Markers
Figures
Expression
Phenotype
Mutations / Transgenics
Human Disease / Model
Sequence Targeting Reagents
Fish
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Orthology
Engineered Foreign Genes
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