TY - JOUR
T1 - Ebola virus VP35 interacts non-covalently with ubiquitin chains to promote viral replication
AU - Rodríguez-Salazar, Carlos A.
AU - van Tol, Sarah
AU - Mailhot, Olivier
AU - Gonzalez-Orozco, Maria
AU - Galdino, Gabriel T.
AU - Warren, Abbey N.
AU - Teruel, Natalia
AU - Behera, Padmanava
AU - Afreen, Kazi Sabrina
AU - Zhang, Lihong
AU - Juelich, Terry L.
AU - Smith, Jennifer K.
AU - Zylber, María Inés
AU - Freiberg, Alexander N.
AU - Najmanovich, Rafael J.
AU - Giraldo, Maria I.
AU - Rajsbaum, Ricardo
N1 - Publisher Copyright:
© 2024 Rodríguez-Salazar et al.
PY - 2024/2
Y1 - 2024/2
N2 - AU Ebolavirus: Pleaseconfirmthatallheadinglevelsarerepresentedcorrectly (EBOV) belongs to a family of highly pathogenic :viruses that cause severe hemorrhagic fever in humans. EBOV replication requires the activity of the viral polymerase complex, which includes the cofactor and Interferon antagonist VP35. We previously showed that the covalent ubiquitination of VP35 promotes virus replication by regulating interactions with the polymerase complex. In addition, VP35 can also interact non-covalently with ubiquitin (Ub); however, the function of this interaction is unknown. Here, we report that VP35 interacts with free (unanchored) K63-linked polyUb chains. Ectopic expression of Isopeptidase T (USP5), which is known to degrade unanchored polyUb chains, reduced VP35 association with Ub and correlated with diminished polymerase activity in a minigenome assay. Using computational methods, we modeled the VP35-Ub non-covalent interacting complex, identified the VP35-Ub interacting surface, and tested mutations to validate the interface. Docking simulations identified chemical compounds that can block VP35-Ub interactions leading to reduced viral polymerase activity. Treatment with the compounds reduced replication of infectious EBOV in cells and in vivo in a mouse model. In conclusion, we identified a novel role of unanchored polyUb in regulating Ebola virus polymerase function and discovered compounds that have promising anti-Ebola virus activity.
AB - AU Ebolavirus: Pleaseconfirmthatallheadinglevelsarerepresentedcorrectly (EBOV) belongs to a family of highly pathogenic :viruses that cause severe hemorrhagic fever in humans. EBOV replication requires the activity of the viral polymerase complex, which includes the cofactor and Interferon antagonist VP35. We previously showed that the covalent ubiquitination of VP35 promotes virus replication by regulating interactions with the polymerase complex. In addition, VP35 can also interact non-covalently with ubiquitin (Ub); however, the function of this interaction is unknown. Here, we report that VP35 interacts with free (unanchored) K63-linked polyUb chains. Ectopic expression of Isopeptidase T (USP5), which is known to degrade unanchored polyUb chains, reduced VP35 association with Ub and correlated with diminished polymerase activity in a minigenome assay. Using computational methods, we modeled the VP35-Ub non-covalent interacting complex, identified the VP35-Ub interacting surface, and tested mutations to validate the interface. Docking simulations identified chemical compounds that can block VP35-Ub interactions leading to reduced viral polymerase activity. Treatment with the compounds reduced replication of infectious EBOV in cells and in vivo in a mouse model. In conclusion, we identified a novel role of unanchored polyUb in regulating Ebola virus polymerase function and discovered compounds that have promising anti-Ebola virus activity.
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U2 - 10.1371/journal.pbio.3002544
DO - 10.1371/journal.pbio.3002544
M3 - Article
C2 - 38422166
AN - SCOPUS:85186389053
SN - 1544-9173
VL - 22
JO - PLoS Biology
JF - PLoS Biology
IS - 2
M1 - e3002544
ER -