A novel, essential trans-splicing protein connects the nematode SL1 snRNP to the CBC-ARS2 complex.
Journal
Nucleic acids research
ISSN: 1362-4962
Titre abrégé: Nucleic Acids Res
Pays: England
ID NLM: 0411011
Informations de publication
Date de publication:
22 07 2022
22 07 2022
Historique:
accepted:
07
06
2022
revised:
01
06
2022
received:
09
07
2021
pubmed:
24
6
2022
medline:
26
7
2022
entrez:
23
6
2022
Statut:
ppublish
Résumé
Spliced leader trans-splicing is essential for gene expression in many eukaryotes. To elucidate the molecular mechanism of this process, we characterise the molecules associated with the Caenorhabditis elegans major spliced leader snRNP (SL1 snRNP), which donates the spliced leader that replaces the 5' untranslated region of most pre-mRNAs. Using a GFP-tagged version of the SL1 snRNP protein SNA-1 created by CRISPR-mediated genome engineering, we immunoprecipitate and identify RNAs and protein components by RIP-Seq and mass spectrometry. This reveals the composition of the SL1 snRNP and identifies associations with spliceosome components PRP-8 and PRP-19. Significantly, we identify a novel, nematode-specific protein required for SL1 trans-splicing, which we designate SNA-3. SNA-3 is an essential, nuclear protein with three NADAR domains whose function is unknown. Mutation of key residues in NADAR domains inactivates the protein, indicating that domain function is required for activity. SNA-3 interacts with the CBC-ARS2 complex and other factors involved in RNA metabolism, including SUT-1 protein, through RNA or protein-mediated contacts revealed by yeast two-hybrid assays, localisation studies and immunoprecipitations. Our data are compatible with a role for SNA-3 in coordinating trans-splicing with target pre-mRNA transcription or in the processing of the Y-branch product of the trans-splicing reaction.
Identifiants
pubmed: 35736244
pii: 6613924
doi: 10.1093/nar/gkac534
pmc: PMC9303266
doi:
Substances chimiques
5' Untranslated Regions
0
Caenorhabditis elegans Proteins
0
Nuclear Proteins
0
Ribonucleoproteins, Small Nuclear
0
RNA, Helminth
0
RNA, Spliced Leader
0
sna-3 protein, C elegans
0
Types de publication
Journal Article
Research Support, N.I.H., Extramural
Research Support, Non-U.S. Gov't
Langues
eng
Sous-ensembles de citation
IM
Pagination
7591-7607Subventions
Organisme : Biotechnology and Biological Sciences Research Council
ID : BB/M010996/1
Pays : United Kingdom
Organisme : NIH HHS
ID : P40 OD010440
Pays : United States
Organisme : Biotechnology and Biological Sciences Research Council
ID : BB/T002859/1
Pays : United Kingdom
Informations de copyright
© The Author(s) 2022. Published by Oxford University Press on behalf of Nucleic Acids Research.
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