Unveiling the role of PUS7-mediated pseudouridylation in host protein interactions specific for the SARS-CoV-2 RNA genome
Giambruno, Roberto (Fondazione Istituto Italiano di Tecnologia (Milà, Itàlia))
Zacco, Elsa (Istituto Italiano di Tecnologia (Gènova, Itàlia))
Ugolini, Camilla (University of Milan)
Vandelli, Andrea (Universitat Autònoma de Barcelona. Departament de Bioquímica i de Biologia Molecular)
Mulroney, Logan (European Molecular Biology Laboratory (Monterotondo, Itàlia))
D'Onghia, Manfredi (Fondazione Istituto Italiano di Tecnologia (Milà, Itàlia))
Giuliani, Bianca (Fondazione Istituto Italiano di Tecnologia (Milà, Itàlia))
Criscuolo, Elena (Vita-Salute San Raffaele University (Milà, Itàlia))
Castelli, Matteo (Vita-Salute San Raffaele University (Milà, Itàlia))
Clementi, Nicola (IRCCS San Raffaele Scientific Institute (Milà, Itàlia))
Clementi, Massimo (IRCCS San Raffaele Scientific Institute (Milà, Itàlia))
Mancini, Nicasio (IRCCS San Raffaele Scientific Institute (Milà, Itàlia))
Bonaldi, Tiziana (University of Milan)
Gustincich, Stefano (Istituto Italiano di Tecnologia (Gènova, Itàlia))
Leonardi, Tommaso (Fondazione Istituto Italiano di Tecnologia (Milà, Itàlia))
Tartaglia, Gian Gaetano (Institució Catalana de Recerca i Estudis Avançats)
Nicassio, Francesco (Istituto Italiano di Tecnologia (Gènova, Itàlia))

Fecha:	2023
Resumen:	Severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2), a positive single-stranded RNA virus, engages in complex interactions with host cell proteins throughout its life cycle. While these interactions enable the host to recognize and inhibit viral replication, they also facilitate essential viral processes such as transcription, translation, and replication. Many aspects of these virus-host interactions remain poorly understood. Here, we employed the cat RAPID algorithm and utilized the RNA-protein interaction detection coupled with mass spectrometry technology to predict and validate the host proteins that specifically bind to the highly structured 5' and 3' terminal regions of the SARS-CoV-2 RNA. Among the interactions identified, we prioritized pseudouridine synthase PUS7, which binds to both ends of the viral RNA. Using nanopore direct RNA sequencing, we discovered that the viral RNA undergoes extensive post-transcriptional modifications. Modified consensus regions for PUS7 were identified at both terminal regions of the SARS-CoV-2 RNA, including one in the viral transcription regulatory sequence leader. Collectively, our findings offer insights into host protein interactions with the SARS-CoV-2 UTRs and highlight the likely significance of pseudouridine synthases and other post-transcriptional modifications in the viral life cycle. This new knowledge enhances our understanding of virus-host dynamics and could inform the development of targeted therapeutic strategies. Using proteomic approaches and in silico predictions, we identified the human host proteins interacting with the terminal regions of SARS-CoV-2 genome. Among these is PUS7, an enzyme converting uridine into pseudouridine. Nanopore sequencing confirmed that viral RNAs contained modified PUS7 consensus sequences, suggesting a role for PUS7 in the virus biology.
Derechos:	Aquest document està subjecte a una llicència d'ús Creative Commons. Es permet la reproducció total o parcial, la distribució, i la comunicació pública de l'obra, sempre que no sigui amb finalitats comercials, i sempre que es reconegui l'autoria de l'obra original. No es permet la creació d'obres derivades.
Lengua:	Anglès
Documento:	Article ; recerca ; Versió publicada
Materia:	MT: RNA/DNA editing ; Protein-rna interactions ; SARS-CoV-2 ; Pseudouridine ; PUS7 ; Proximity biotinylation ; Cat RAPID ; Nanocompore ; Nanopore direct-RNA sequencing ; RNA-binding proteins
Publicado en:	Molecular Therapy. Nucleic Acids, Vol. 34 (october 2023) , ISSN 2162-2531

DOI: 10.1016/j.omtn.2023.102052
PMID: 38028201

17 p, 2.8 MB

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