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  1. Article ; Online: Identifying cellular RNA-binding proteins during infection uncovers a role for MKRN2 in influenza mRNA trafficking.

    Bonazza, Stefano / Coutts, Hannah Leigh / Sukumar, Swathi / Turkington, Hannah Louise / Courtney, David Gary

    PLoS pathogens

    2024  Volume 20, Issue 5, Page(s) e1012231

    Abstract: Utilisation of RNA-binding proteins (RBPs) is an important aspect of post-transcriptional regulation of viral RNA. Viruses such as influenza A viruses (IAV) interact with RBPs to regulate processes including splicing, nuclear export and trafficking, ... ...

    Abstract Utilisation of RNA-binding proteins (RBPs) is an important aspect of post-transcriptional regulation of viral RNA. Viruses such as influenza A viruses (IAV) interact with RBPs to regulate processes including splicing, nuclear export and trafficking, while also encoding RBPs within their genomes, such as NP and NS1. But with almost 1000 RBPs encoded within the human genome it is still unclear what role, if any, many of these proteins play during viral replication. Using the RNA interactome capture (RIC) technique, we isolated RBPs from IAV infected cells to unravel the RBPome of mRNAs from IAV infected human cells. This led to the identification of one particular RBP, MKRN2, that associates with and positively regulates IAV mRNA. Through further validation, we determined that MKRN2 is involved in the nuclear-cytoplasmic trafficking of IAV mRNA potentially through an association with the RNA export mediator GLE1. In the absence of MKRN2, IAV mRNAs accumulate in the nucleus of infected cells, which may lead to their degradation by the nuclear RNA exosome complex. MKRN2, therefore, appears to be required for the efficient nuclear export of IAV mRNAs in human cells.
    Language English
    Publishing date 2024-05-16
    Publishing country United States
    Document type Journal Article
    ZDB-ID 2205412-1
    ISSN 1553-7374 ; 1553-7374
    ISSN (online) 1553-7374
    ISSN 1553-7374
    DOI 10.1371/journal.ppat.1012231
    Database MEDical Literature Analysis and Retrieval System OnLINE

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  2. Article ; Online: Human cytomegalovirus forms phase-separated compartments at viral genomes to facilitate viral replication.

    Caragliano, Enrico / Bonazza, Stefano / Frascaroli, Giada / Tang, Jiajia / Soh, Timothy K / Grünewald, Kay / Bosse, Jens B / Brune, Wolfram

    Cell reports

    2022  Volume 38, Issue 10, Page(s) 110469

    Abstract: Human cytomegalovirus (HCMV) replicates its DNA genome in specialized replication compartments (RCs) in the host cell nucleus. These membrane-less organelles originate as spherical structures and grow in size over time. However, the mechanism of RC ... ...

    Abstract Human cytomegalovirus (HCMV) replicates its DNA genome in specialized replication compartments (RCs) in the host cell nucleus. These membrane-less organelles originate as spherical structures and grow in size over time. However, the mechanism of RC biogenesis has remained understudied. Using live-cell imaging and photo-oligomerization, we show that a central component of RCs, the UL112-113 proteins, undergo liquid-liquid phase separation (LLPS) to form RCs in the nucleus. We show that the self-interacting domain and large intrinsically disordered regions of UL112-113 are required for LLPS. Importantly, viral DNA induces local clustering of these proteins and lowers the threshold for phase separation. The formation of phase-separated compartments around viral genomes is necessary to recruit the viral DNA polymerase for viral genome replication. Thus, HCMV uses its UL112-113 proteins to generate RCs around viral genomes by LLPS to ensure the formation of a pro-replicative environment.
    MeSH term(s) Cell Nucleus/metabolism ; Cytomegalovirus/genetics ; Cytomegalovirus/metabolism ; DNA, Viral/genetics ; DNA, Viral/metabolism ; Genome, Viral ; Humans ; Viral Proteins/metabolism ; Virus Replication
    Chemical Substances DNA, Viral ; Viral Proteins
    Language English
    Publishing date 2022-03-19
    Publishing country United States
    Document type Journal Article ; Research Support, Non-U.S. Gov't
    ZDB-ID 2649101-1
    ISSN 2211-1247 ; 2211-1247
    ISSN (online) 2211-1247
    ISSN 2211-1247
    DOI 10.1016/j.celrep.2022.110469
    Database MEDical Literature Analysis and Retrieval System OnLINE

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  3. Article ; Online: The RBPome of influenza A virus NP-mRNA reveals a role for TDP-43 in viral replication.

    Dupont, Maud / Krischuns, Tim / Gianetto, Quentin Giai / Paisant, Sylvain / Bonazza, Stefano / Brault, Jean-Baptiste / Douché, Thibaut / Arragain, Benoît / Florez-Prada, Alberto / Perez-Perri, Joel I / Hentze, Matthias W / Cusack, Stephen / Matondo, Mariette / Isel, Catherine / Courtney, David G / Naffakh, Nadia

    Nucleic acids research

    2024  

    Abstract: Genome-wide approaches have significantly advanced our knowledge of the repertoire of RNA-binding proteins (RBPs) that associate with cellular polyadenylated mRNAs within eukaryotic cells. Recent studies focusing on the RBP interactomes of viral mRNAs, ... ...

    Abstract Genome-wide approaches have significantly advanced our knowledge of the repertoire of RNA-binding proteins (RBPs) that associate with cellular polyadenylated mRNAs within eukaryotic cells. Recent studies focusing on the RBP interactomes of viral mRNAs, notably SARS-Cov-2, have revealed both similarities and differences between the RBP profiles of viral and cellular mRNAs. However, the RBPome of influenza virus mRNAs remains unexplored. Herein, we identify RBPs that associate with the viral mRNA encoding the nucleoprotein (NP) of an influenza A virus. Focusing on TDP-43, we show that it binds several influenza mRNAs beyond the NP-mRNA, and that its depletion results in lower levels of viral mRNAs and proteins within infected cells, and a decreased yield of infectious viral particles. We provide evidence that the viral polymerase recruits TDP-43 onto viral mRNAs through a direct interaction with the disordered C-terminal domain of TDP-43. Notably, other RBPs found to be associated with influenza virus mRNAs also interact with the viral polymerase, which points to a role of the polymerase in orchestrating the assembly of viral messenger ribonucleoproteins.
    Language English
    Publishing date 2024-04-30
    Publishing country England
    Document type Journal Article
    ZDB-ID 186809-3
    ISSN 1362-4962 ; 1362-4954 ; 0301-5610 ; 0305-1048
    ISSN (online) 1362-4962 ; 1362-4954
    ISSN 0301-5610 ; 0305-1048
    DOI 10.1093/nar/gkae291
    Database MEDical Literature Analysis and Retrieval System OnLINE

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