6Z9U

Crystal structure of a TSEN15-34 heterodimer.

Summary for 6Z9U

• Entry DOI: 10.2210/pdb6z9u/pdb
• Descriptor: tRNA-splicing endonuclease subunit Sen34, tRNA-splicing endonuclease subunit Sen15, GLYCEROL, ... (4 entities in total)
• Functional Keywords: trna splicing endonuclease, tsen, precursor trna, pre-trna processing, neurodegenerative disorders, pontocerebellar hypoplasia, splicing
• Biological source: Homo sapiens (Human); More
• Total number of polymer chains: 4
• Total formula weight: 61201.40

Authors

Trowitzsch, S.,Sekulovski, S. (deposition date: 2020-06-04, release date: 2021-06-30, Last modification date: 2024-01-24)

Primary citation

Sekulovski, S.,Devant, P.,Panizza, S.,Gogakos, T.,Pitiriciu, A.,Heitmeier, K.,Ramsay, E.P.,Barth, M.,Schmidt, C.,Tuschl, T.,Baas, F.,Weitzer, S.,Martinez, J.,Trowitzsch, S.
Assembly defects of human tRNA splicing endonuclease contribute to impaired pre-tRNA processing in pontocerebellar hypoplasia.
Nat Commun, 12:5610-5610, 2021

PubMed Abstract: 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.

PubMed: 34584079
DOI: 10.1038/s41467-021-25870-3

Experimental method

X-RAY DIFFRACTION (2.10001777636 Å)