TY  - INPR
A1  - Sekulovski, Samoil
A1  - Devant, Pascal
A1  - Panizza, Silvia
A1  - Gogakos, Tasos
A1  - Pitiriciu, Anda
A1  - Heitmeier, Katharina
A1  - Ramsay, Ewan Phillip
A1  - Barth, Marie
A1  - Schmidt, Carla
A1  - Weitzer, Stefan
A1  - Tuschl, Thomas
A1  - Baas, Frank
A1  - Martinez, Javier
A1  - Trowitzsch, Simon
T1  - Assembly defects of the human tRNA splicing endonuclease contribute to impaired pre-tRNA processing in pontocerebellar hypoplasia
T2  - bioRxiv
N2  - Introns of human transfer RNA precursors (pre-tRNAs) are excised by the tRNA splicing endonuclease TSEN in complex with the RNA kinase CLP1. Mutations in TSEN/CLP1 occur in patients with pontocerebellar hypoplasia (PCH), however, their role in the disease is unclear. Here, we show that intron excision is catalyzed by tetrameric TSEN assembled from inactive heterodimers independently of CLP1. Splice site recognition involves the mature domain and the anticodon-intron base pair of pre-tRNAs. The 2.1-Å resolution X-ray crystal structure of a TSEN15–34 heterodimer and differential scanning fluorimetry analyses show that PCH mutations cause thermal destabilization. While endonuclease activity in recombinant mutant TSEN is unaltered, we observe assembly defects and reduced pre-tRNA cleavage activity resulting in an imbalanced pre-tRNA pool in PCH patient-derived fibroblasts. Our work defines the molecular principles of intron excision in humans and provides evidence that modulation of TSEN stability may contribute to PCH phenotypes.
Y1  - 2020
UR  - http://publikationen.ub.uni-frankfurt.de/frontdoor/index/index/docId/72809
UR  - https://nbn-resolving.org/urn:nbn:de:hebis:30:3-728096
IS  - 2020.08.03.234229
ER  -