Structural basis for pre-tRNA recognition and processing by the human tRNA splicing endonuclease complex.

Cassandra K Hayne; Kevin John U Butay; Zachary D Stewart; Juno M Krahn; Lalith Perera; Jason G Williams; Robert M Petrovitch; Leesa J Deterding; A Gregory Matera; Mario J Borgnia; Robin E Stanley

doi:10.1038/s41594-023-00991-z

PubMed · 37231153

Structural basis for pre-tRNA recognition and processing by the human tRNA splicing endonuclease complex.

Cassandra K Hayne·Kevin John U Butay·Zachary D Stewart·Juno M Krahn·Lalith Perera·Jason G Williams·Robert M Petrovitch·Leesa J Deterding·A Gregory Matera·Mario J Borgnia·Robin E Stanley

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Abstract

Throughout bacteria, archaea and eukarya, certain tRNA transcripts contain introns. Pre-tRNAs with introns require splicing to form the mature anticodon stem loop. In eukaryotes, tRNA splicing is initiated by the heterotetrameric tRNA splicing endonuclease (TSEN) complex. All TSEN subunits are essential, and mutations within the complex are associated with a family of neurodevelopmental disorders known as pontocerebellar hypoplasia (PCH). Here, we report cryo-electron microscopy structures of the human TSEN-pre-tRNA complex. These structures reveal the overall architecture of the complex and the extensive tRNA binding interfaces. The structures share homology with archaeal TSENs but contain additional features important for pre-tRNA recognition. The TSEN54 subunit functions as a pivotal scaffold for the pre-tRNA and the two endonuclease subunits. Finally, the TSEN structures enable visualization of the molecular environments of PCH-causing missense mutations, providing insight into the mechanism of pre-tRNA splicing and PCH.

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Humans RNA Precursors Cryoelectron Microscopy Endoribonucleases RNA Splicing Introns RNA, Transfer Archaea Eukaryota Nucleic Acid Conformation Journal Article Research Support, N.I.H., Extramural Research Support, N.I.H., Intramural

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Cassandra K Hayne, Kevin John U Butay, Zachary D Stewart, Juno M Krahn, Lalith Perera, Jason G Williams, Robert M Petrovitch, Leesa J Deterding, A Gregory Matera, Mario J Borgnia, Robin E Stanley. 2023-05-25. Structural basis for pre-tRNA recognition and processing by the human tRNA splicing endonuclease complex.. https://doi.org/10.1038/s41594-023-00991-z

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