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Identification of functionally important domains of human cytomegalovirus gO that act after trimer binding to receptors

Human cytomegalovirus (HCMV) entry involves trimer (gH/gL/gO) that interacts with PDGFRα in fibroblasts. Entry into epithelial and endothelial cells requires trimer, which binds unidentified receptors, and pentamer (gH/gL/UL128-131), which binds neuropilin-2. To identify functionally important domai...

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Autores principales: Chin, Andrea, Liu, Jing, Jardetzky, Theodore, Johnson, David C., Vanarsdall, Adam
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Public Library of Science 2022
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC9032346/
https://www.ncbi.nlm.nih.gov/pubmed/35452493
http://dx.doi.org/10.1371/journal.ppat.1010452
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author Chin, Andrea
Liu, Jing
Jardetzky, Theodore
Johnson, David C.
Vanarsdall, Adam
author_facet Chin, Andrea
Liu, Jing
Jardetzky, Theodore
Johnson, David C.
Vanarsdall, Adam
author_sort Chin, Andrea
collection PubMed
description Human cytomegalovirus (HCMV) entry involves trimer (gH/gL/gO) that interacts with PDGFRα in fibroblasts. Entry into epithelial and endothelial cells requires trimer, which binds unidentified receptors, and pentamer (gH/gL/UL128-131), which binds neuropilin-2. To identify functionally important domains in trimer, we screened an overlapping 20-mer gO peptide library and identified two sets of peptides: 19/20 (a.a. 235–267) and 32/33 (a.a. 404–436) that could block virus entry. Soluble trimer containing wild type gO blocked HCMV entry, whereas soluble trimers with the 19/20 or 32/33 sequences mutated did not block entry. Interestingly, the mutant trimers retained the capacity to bind to cellular receptors including PDGFRα. Peptide 19/20 and 32/33 sequences formed a lobe extending from the surface of gO and an adjacent concave structure, respectively. Neither of these sets of sequences contacted PDGFRα. Instead, our data support a model in which the 19/20 and 32/33 trimer sequences function downstream of receptor binding, e.g. trafficking of HCMV into endosomes or binding to gB for entry fusion. We also screened for peptides that bound antibodies (Abs) in human sera, observing that peptides 20 and 26 bound Abs. These peptides engendered neutralizing Abs (NAbs) after immunization of rabbits and could pull out NAbs from human sera. Peptides 20 and 26 sequences represent the first NAb epitopes identified in trimer. These studies describe two important surfaces on gO defined by: i) peptides 19/20 and 32/33, which apparently act downstream of receptor binding and ii) peptide 26 that interacts with PDGFRα. Both these surfaces are targets of NAbs.
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spelling pubmed-90323462022-04-23 Identification of functionally important domains of human cytomegalovirus gO that act after trimer binding to receptors Chin, Andrea Liu, Jing Jardetzky, Theodore Johnson, David C. Vanarsdall, Adam PLoS Pathog Research Article Human cytomegalovirus (HCMV) entry involves trimer (gH/gL/gO) that interacts with PDGFRα in fibroblasts. Entry into epithelial and endothelial cells requires trimer, which binds unidentified receptors, and pentamer (gH/gL/UL128-131), which binds neuropilin-2. To identify functionally important domains in trimer, we screened an overlapping 20-mer gO peptide library and identified two sets of peptides: 19/20 (a.a. 235–267) and 32/33 (a.a. 404–436) that could block virus entry. Soluble trimer containing wild type gO blocked HCMV entry, whereas soluble trimers with the 19/20 or 32/33 sequences mutated did not block entry. Interestingly, the mutant trimers retained the capacity to bind to cellular receptors including PDGFRα. Peptide 19/20 and 32/33 sequences formed a lobe extending from the surface of gO and an adjacent concave structure, respectively. Neither of these sets of sequences contacted PDGFRα. Instead, our data support a model in which the 19/20 and 32/33 trimer sequences function downstream of receptor binding, e.g. trafficking of HCMV into endosomes or binding to gB for entry fusion. We also screened for peptides that bound antibodies (Abs) in human sera, observing that peptides 20 and 26 bound Abs. These peptides engendered neutralizing Abs (NAbs) after immunization of rabbits and could pull out NAbs from human sera. Peptides 20 and 26 sequences represent the first NAb epitopes identified in trimer. These studies describe two important surfaces on gO defined by: i) peptides 19/20 and 32/33, which apparently act downstream of receptor binding and ii) peptide 26 that interacts with PDGFRα. Both these surfaces are targets of NAbs. Public Library of Science 2022-04-22 /pmc/articles/PMC9032346/ /pubmed/35452493 http://dx.doi.org/10.1371/journal.ppat.1010452 Text en © 2022 Chin et al https://creativecommons.org/licenses/by/4.0/This is an open access article distributed under the terms of the Creative Commons Attribution License (https://creativecommons.org/licenses/by/4.0/) , which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited.
spellingShingle Research Article
Chin, Andrea
Liu, Jing
Jardetzky, Theodore
Johnson, David C.
Vanarsdall, Adam
Identification of functionally important domains of human cytomegalovirus gO that act after trimer binding to receptors
title Identification of functionally important domains of human cytomegalovirus gO that act after trimer binding to receptors
title_full Identification of functionally important domains of human cytomegalovirus gO that act after trimer binding to receptors
title_fullStr Identification of functionally important domains of human cytomegalovirus gO that act after trimer binding to receptors
title_full_unstemmed Identification of functionally important domains of human cytomegalovirus gO that act after trimer binding to receptors
title_short Identification of functionally important domains of human cytomegalovirus gO that act after trimer binding to receptors
title_sort identification of functionally important domains of human cytomegalovirus go that act after trimer binding to receptors
topic Research Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC9032346/
https://www.ncbi.nlm.nih.gov/pubmed/35452493
http://dx.doi.org/10.1371/journal.ppat.1010452
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