Giardia lamblia Hsp90 pre-mRNAs undergo self-splicing to generate mature RNA in an in vitro trans-splicing reaction.
G. lamblia
RNA repair
RNA splicing
heat shock protein 90
molecular chaperone
post-transcriptional regulation
Journal
FEBS letters
ISSN: 1873-3468
Titre abrégé: FEBS Lett
Pays: England
ID NLM: 0155157
Informations de publication
Date de publication:
02 2019
02 2019
Historique:
received:
23
11
2018
accepted:
26
12
2018
pubmed:
4
1
2019
medline:
18
10
2019
entrez:
4
1
2019
Statut:
ppublish
Résumé
We have previously shown that the Heat Shock Protein 90 (Hsp90) gene in G. lamblia is expressed from two ORFs localized 777 kb apart. The pre-mRNAs transcribed from these ORFs are stitched by a trans-splicing mechanism. Here, we provide mechanistic details of this process by reconstituting the reaction using in vitro synthesized pre-mRNA substrates. Using RT-PCR, northern blot and nanostring technology, we demonstrate that the in vitro synthesized pre-mRNAs have the capability to self-splice in the absence of nuclear proteins. Inhibition of the trans-splicing reaction using a ssDNA oligo corresponding to a 26-nucleotide complementary sequence confirmed their role in juxtapositioning the pre-mRNA substrates during the self-splicing reaction. Our study provides the first example of a self catalysed, trans-splicing reaction in eukaryotes.
Identifiants
pubmed: 30604864
doi: 10.1002/1873-3468.13324
doi:
Substances chimiques
DNA, Single-Stranded
0
HSP90 Heat-Shock Proteins
0
Protozoan Proteins
0
RNA Precursors
0
Types de publication
Journal Article
Research Support, Non-U.S. Gov't
Langues
eng
Sous-ensembles de citation
IM
Pagination
433-442Informations de copyright
© 2019 Federation of European Biochemical Societies.