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Tryptophan Residues Are Critical for Portal Protein Assembly and Incorporation in Bacteriophage P22
The oligomerization and incorporation of the bacteriophage P22 portal protein complex into procapsids (PCs) depends upon an interaction with scaffolding protein, but the region of the portal protein that interacts with scaffolding protein has not been defined. In herpes simplex virus 1 (HSV-1), cons...
Autores principales: | , , , , , , , , |
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Formato: | Online Artículo Texto |
Lenguaje: | English |
Publicado: |
MDPI
2022
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Materias: | |
Acceso en línea: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC9320234/ https://www.ncbi.nlm.nih.gov/pubmed/35891382 http://dx.doi.org/10.3390/v14071400 |
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author | Woodbury, Brianna M. Motwani, Tina Leroux, Makayla N. Barnes, Lauren F. Lyktey, Nicholas A. Banerjee, Sanchari Dedeo, Corynne L. Jarrold, Martin F. Teschke, Carolyn M. |
author_facet | Woodbury, Brianna M. Motwani, Tina Leroux, Makayla N. Barnes, Lauren F. Lyktey, Nicholas A. Banerjee, Sanchari Dedeo, Corynne L. Jarrold, Martin F. Teschke, Carolyn M. |
author_sort | Woodbury, Brianna M. |
collection | PubMed |
description | The oligomerization and incorporation of the bacteriophage P22 portal protein complex into procapsids (PCs) depends upon an interaction with scaffolding protein, but the region of the portal protein that interacts with scaffolding protein has not been defined. In herpes simplex virus 1 (HSV-1), conserved tryptophan residues located in the wing domain are required for portal-scaffolding protein interactions. In this study, tryptophan residues (W) present at positions 41, 44, 207 and 211 within the wing domain of the bacteriophage P22 portal protein were mutated to both conserved and non-conserved amino acids. Substitutions at each of these positions were shown to impair portal function in vivo, resulting in a lethal phenotype by complementation. The alanine substitutions caused the most severe defects and were thus further characterized. An analysis of infected cell lysates for the W to A mutants revealed that all the portal protein variants except W211A, which has a temperature-sensitive incorporation defect, were successfully recruited into procapsids. By charge detection mass spectrometry, all W to A mutant portal proteins were shown to form stable dodecameric rings except the variant W41A, which dissociated readily to monomers. Together, these results suggest that for P22 conserved tryptophan, residues in the wing domain of the portal protein play key roles in portal protein oligomerization and incorporation into procapsids, ultimately affecting the functionality of the portal protein at specific stages of virus assembly. |
format | Online Article Text |
id | pubmed-9320234 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2022 |
publisher | MDPI |
record_format | MEDLINE/PubMed |
spelling | pubmed-93202342022-07-27 Tryptophan Residues Are Critical for Portal Protein Assembly and Incorporation in Bacteriophage P22 Woodbury, Brianna M. Motwani, Tina Leroux, Makayla N. Barnes, Lauren F. Lyktey, Nicholas A. Banerjee, Sanchari Dedeo, Corynne L. Jarrold, Martin F. Teschke, Carolyn M. Viruses Article The oligomerization and incorporation of the bacteriophage P22 portal protein complex into procapsids (PCs) depends upon an interaction with scaffolding protein, but the region of the portal protein that interacts with scaffolding protein has not been defined. In herpes simplex virus 1 (HSV-1), conserved tryptophan residues located in the wing domain are required for portal-scaffolding protein interactions. In this study, tryptophan residues (W) present at positions 41, 44, 207 and 211 within the wing domain of the bacteriophage P22 portal protein were mutated to both conserved and non-conserved amino acids. Substitutions at each of these positions were shown to impair portal function in vivo, resulting in a lethal phenotype by complementation. The alanine substitutions caused the most severe defects and were thus further characterized. An analysis of infected cell lysates for the W to A mutants revealed that all the portal protein variants except W211A, which has a temperature-sensitive incorporation defect, were successfully recruited into procapsids. By charge detection mass spectrometry, all W to A mutant portal proteins were shown to form stable dodecameric rings except the variant W41A, which dissociated readily to monomers. Together, these results suggest that for P22 conserved tryptophan, residues in the wing domain of the portal protein play key roles in portal protein oligomerization and incorporation into procapsids, ultimately affecting the functionality of the portal protein at specific stages of virus assembly. MDPI 2022-06-27 /pmc/articles/PMC9320234/ /pubmed/35891382 http://dx.doi.org/10.3390/v14071400 Text en © 2022 by the authors. https://creativecommons.org/licenses/by/4.0/Licensee MDPI, Basel, Switzerland. This article is an open access article distributed under the terms and conditions of the Creative Commons Attribution (CC BY) license (https://creativecommons.org/licenses/by/4.0/). |
spellingShingle | Article Woodbury, Brianna M. Motwani, Tina Leroux, Makayla N. Barnes, Lauren F. Lyktey, Nicholas A. Banerjee, Sanchari Dedeo, Corynne L. Jarrold, Martin F. Teschke, Carolyn M. Tryptophan Residues Are Critical for Portal Protein Assembly and Incorporation in Bacteriophage P22 |
title | Tryptophan Residues Are Critical for Portal Protein Assembly and Incorporation in Bacteriophage P22 |
title_full | Tryptophan Residues Are Critical for Portal Protein Assembly and Incorporation in Bacteriophage P22 |
title_fullStr | Tryptophan Residues Are Critical for Portal Protein Assembly and Incorporation in Bacteriophage P22 |
title_full_unstemmed | Tryptophan Residues Are Critical for Portal Protein Assembly and Incorporation in Bacteriophage P22 |
title_short | Tryptophan Residues Are Critical for Portal Protein Assembly and Incorporation in Bacteriophage P22 |
title_sort | tryptophan residues are critical for portal protein assembly and incorporation in bacteriophage p22 |
topic | Article |
url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC9320234/ https://www.ncbi.nlm.nih.gov/pubmed/35891382 http://dx.doi.org/10.3390/v14071400 |
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