TY - JOUR
T1 - DHX15-independent roles for TFIP11 in U6 snRNA modification, U4/U6.U5 tri-snRNP assembly and pre-mRNA splicing fidelity
AU - Duchemin, Amandine
AU - O'Grady, Tina
AU - Hanache, Sarah
AU - Mereau, Agnès
AU - Thiry, Marc
AU - Wacheul, Ludivine
AU - Michaux, Catherine
AU - Perpete, Eric
AU - Hervouet, Eric
AU - Peixoto, Paul
AU - Ernst, Felix GM
AU - Audic, Yann
AU - Dequiedt, Franck
AU - Lafontaine, Denis LJ
AU - Mottet, Denis
N1 - Funding Information:
The authors thank the GIGA Cell Imaging and Flow Cytometry and the GIGA Tran-scriptomic core facilities for technical assistance. We also thank the GIGA scientific illustrator, Adeline Deward, for designing and formatting figures and Rümeyza Bascetin (RNA Molecular Biology, ULB, Belgium) for her advice in microscopy. We also thank the Genouest bioinformatic platform at the IGDR (Rennes, France) and the BRIGHTcore facility at the Université Libre de Bruxelles (ULB, Belgium) for providing computing resources for iCLIP-Seq and RiboMethSeq experiments, respectively. Computational resources have been provided by the Consortium des Équipements de Calcul Intensif (CÉCI), funded by the National Fund for Scientific Research (FNRS) under Grant No. 2.5020.11 and by the Walloon Region. This work was supported by grants from the FNRS–Belgium (CDR Grant No. J003618F), TELEVIE, Fonds Léon Frédéricq and Fonds Spéciaux de la Recherche de l’Université de Liège (ULiege). A.D. is FNRS-FRIA fellow. T.O’G. is FNRS Postdoctoral Researcher. S.H. is FNRS-TELEVIE fellow. E.P. and D.L.J.L. are FNRS Senior Research Associates. C.M. and D.M. are FNRS Research Associates.
Publisher Copyright:
© 2021, The Author(s).
PY - 2021/12
Y1 - 2021/12
N2 - The U6 snRNA, the core catalytic component of the spliceosome, is extensively modified post-transcriptionally, with 2’-O-methylation being most common. However, how U6 2’-O-methylation is regulated remains largely unknown. Here we report that TFIP11, the human homolog of the yeast spliceosome disassembly factor Ntr1, localizes to nucleoli and Cajal Bodies and is essential for the 2’-O-methylation of U6. Mechanistically, we demonstrate that TFIP11 knockdown reduces the association of U6 snRNA with fibrillarin and associated snoRNAs, therefore altering U6 2′-O-methylation. We show U6 snRNA hypomethylation is associated with changes in assembly of the U4/U6.U5 tri-snRNP leading to defects in spliceosome assembly and alterations in splicing fidelity. Strikingly, this function of TFIP11 is independent of the RNA helicase DHX15, its known partner in yeast. In sum, our study demonstrates an unrecognized function for TFIP11 in U6 snRNP modification and U4/U6.U5 tri-snRNP assembly, identifying TFIP11 as a critical spliceosome assembly regulator.
AB - The U6 snRNA, the core catalytic component of the spliceosome, is extensively modified post-transcriptionally, with 2’-O-methylation being most common. However, how U6 2’-O-methylation is regulated remains largely unknown. Here we report that TFIP11, the human homolog of the yeast spliceosome disassembly factor Ntr1, localizes to nucleoli and Cajal Bodies and is essential for the 2’-O-methylation of U6. Mechanistically, we demonstrate that TFIP11 knockdown reduces the association of U6 snRNA with fibrillarin and associated snoRNAs, therefore altering U6 2′-O-methylation. We show U6 snRNA hypomethylation is associated with changes in assembly of the U4/U6.U5 tri-snRNP leading to defects in spliceosome assembly and alterations in splicing fidelity. Strikingly, this function of TFIP11 is independent of the RNA helicase DHX15, its known partner in yeast. In sum, our study demonstrates an unrecognized function for TFIP11 in U6 snRNP modification and U4/U6.U5 tri-snRNP assembly, identifying TFIP11 as a critical spliceosome assembly regulator.
UR - http://www.scopus.com/inward/record.url?scp=85119302906&partnerID=8YFLogxK
U2 - 10.1038/s41467-021-26932-2
DO - 10.1038/s41467-021-26932-2
M3 - Article
SN - 2041-1723
VL - 12
JO - Nature Communications
JF - Nature Communications
IS - 1
M1 - 6648
ER -