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Mass Spectral Analyses of Salmonella Myovirus SPN3US Reveal Conserved and Divergent Themes in Proteolytic Maturation of Large Icosahedral Capsids

Salmonella myovirus SPN3US has a T = 27 capsid composed of >50 different gene products, including many that are packaged along with the 240 kb genome and ejected into the host cell. Recently, we showed that an essential phage-encoded prohead protease gp245 is responsible for cleavage of proteins...

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Autores principales: Scheuch, Aaron, Moran, Sara A. M., Faraone, Julia N., Unwin, Sophia R., Vu, Gialinh, Benítez, Andrea Denisse, Mohd Redzuan, Nurul Humaira, Molleur, Dana, Pardo, Sammy, Weintraub, Susan T., Thomas, Julie A.
Formato: Online Artículo Texto
Lenguaje:English
Publicado: MDPI 2023
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC10052503/
https://www.ncbi.nlm.nih.gov/pubmed/36992431
http://dx.doi.org/10.3390/v15030723
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author Scheuch, Aaron
Moran, Sara A. M.
Faraone, Julia N.
Unwin, Sophia R.
Vu, Gialinh
Benítez, Andrea Denisse
Mohd Redzuan, Nurul Humaira
Molleur, Dana
Pardo, Sammy
Weintraub, Susan T.
Thomas, Julie A.
author_facet Scheuch, Aaron
Moran, Sara A. M.
Faraone, Julia N.
Unwin, Sophia R.
Vu, Gialinh
Benítez, Andrea Denisse
Mohd Redzuan, Nurul Humaira
Molleur, Dana
Pardo, Sammy
Weintraub, Susan T.
Thomas, Julie A.
author_sort Scheuch, Aaron
collection PubMed
description Salmonella myovirus SPN3US has a T = 27 capsid composed of >50 different gene products, including many that are packaged along with the 240 kb genome and ejected into the host cell. Recently, we showed that an essential phage-encoded prohead protease gp245 is responsible for cleavage of proteins during SPN3US head assembly. This proteolytic maturation step induces major changes in precursor head particles, enabling them to expand and undergo genome packaging. To comprehensively define the composition of the mature SPN3US head and elucidate how it is modified by proteolysis during assembly, we conducted tandem mass spectrometry analysis of purified virions and tailless heads. Fourteen protease cleavage sites were identified in nine proteins, including eight sites not previously identified in head proteins in vivo. Among these was the maturation cleavage site of gp245 which was identical to the autocleavage site we had previously identified in purified recombinant gp245. Our findings underscore the value of employing multiple mass spectrometry-based experimental strategies as a way to enhance the detection of head protein cleavage sites in tailed phages. In addition, our results have identified a conserved set of head proteins in related giant phages that are similarly cleaved by their respective prohead proteases, suggesting that these proteins have important roles in governing the formation and function of large icosahedral capsids.
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spelling pubmed-100525032023-03-30 Mass Spectral Analyses of Salmonella Myovirus SPN3US Reveal Conserved and Divergent Themes in Proteolytic Maturation of Large Icosahedral Capsids Scheuch, Aaron Moran, Sara A. M. Faraone, Julia N. Unwin, Sophia R. Vu, Gialinh Benítez, Andrea Denisse Mohd Redzuan, Nurul Humaira Molleur, Dana Pardo, Sammy Weintraub, Susan T. Thomas, Julie A. Viruses Article Salmonella myovirus SPN3US has a T = 27 capsid composed of >50 different gene products, including many that are packaged along with the 240 kb genome and ejected into the host cell. Recently, we showed that an essential phage-encoded prohead protease gp245 is responsible for cleavage of proteins during SPN3US head assembly. This proteolytic maturation step induces major changes in precursor head particles, enabling them to expand and undergo genome packaging. To comprehensively define the composition of the mature SPN3US head and elucidate how it is modified by proteolysis during assembly, we conducted tandem mass spectrometry analysis of purified virions and tailless heads. Fourteen protease cleavage sites were identified in nine proteins, including eight sites not previously identified in head proteins in vivo. Among these was the maturation cleavage site of gp245 which was identical to the autocleavage site we had previously identified in purified recombinant gp245. Our findings underscore the value of employing multiple mass spectrometry-based experimental strategies as a way to enhance the detection of head protein cleavage sites in tailed phages. In addition, our results have identified a conserved set of head proteins in related giant phages that are similarly cleaved by their respective prohead proteases, suggesting that these proteins have important roles in governing the formation and function of large icosahedral capsids. MDPI 2023-03-10 /pmc/articles/PMC10052503/ /pubmed/36992431 http://dx.doi.org/10.3390/v15030723 Text en © 2023 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
Scheuch, Aaron
Moran, Sara A. M.
Faraone, Julia N.
Unwin, Sophia R.
Vu, Gialinh
Benítez, Andrea Denisse
Mohd Redzuan, Nurul Humaira
Molleur, Dana
Pardo, Sammy
Weintraub, Susan T.
Thomas, Julie A.
Mass Spectral Analyses of Salmonella Myovirus SPN3US Reveal Conserved and Divergent Themes in Proteolytic Maturation of Large Icosahedral Capsids
title Mass Spectral Analyses of Salmonella Myovirus SPN3US Reveal Conserved and Divergent Themes in Proteolytic Maturation of Large Icosahedral Capsids
title_full Mass Spectral Analyses of Salmonella Myovirus SPN3US Reveal Conserved and Divergent Themes in Proteolytic Maturation of Large Icosahedral Capsids
title_fullStr Mass Spectral Analyses of Salmonella Myovirus SPN3US Reveal Conserved and Divergent Themes in Proteolytic Maturation of Large Icosahedral Capsids
title_full_unstemmed Mass Spectral Analyses of Salmonella Myovirus SPN3US Reveal Conserved and Divergent Themes in Proteolytic Maturation of Large Icosahedral Capsids
title_short Mass Spectral Analyses of Salmonella Myovirus SPN3US Reveal Conserved and Divergent Themes in Proteolytic Maturation of Large Icosahedral Capsids
title_sort mass spectral analyses of salmonella myovirus spn3us reveal conserved and divergent themes in proteolytic maturation of large icosahedral capsids
topic Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC10052503/
https://www.ncbi.nlm.nih.gov/pubmed/36992431
http://dx.doi.org/10.3390/v15030723
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