Aspartyl Protease 5 Matures Dense Granule Proteins That Reside at the Host-Parasite Interface in <named-content content-type="genus-species">Toxoplasma gondii</named-content>

ABSTRACT Toxoplasma gondii infects approximately 30% of the world’s population, causing disease primarily during pregnancy and in individuals with weakened immune systems. Toxoplasma secretes and exports effector proteins that modulate the host during infection, and several of these proteins are pro...

Descripción completa

Guardado en:

Detalles Bibliográficos
Autores principales:	Michael J. Coffey, Laura F. Dagley, Simona Seizova, Eugene A. Kapp, Giuseppe Infusini, David S. Roos, Justin A. Boddey, Andrew I. Webb, Christopher J. Tonkin
Formato:	article
Lenguaje:	EN
Publicado:	American Society for Microbiology 2018
Materias:	effector proteomics Toxoplasma virulence Microbiology QR1-502
Acceso en línea:	https://doaj.org/article/ea7a3291199047328178731b63d09cfa
Etiquetas:	Agregar Etiqueta Sin Etiquetas, Sea el primero en etiquetar este registro!

id	oai:doaj.org-article:ea7a3291199047328178731b63d09cfa
record_format	dspace
spelling	oai:doaj.org-article:ea7a3291199047328178731b63d09cfa2021-11-15T15:58:21ZAspartyl Protease 5 Matures Dense Granule Proteins That Reside at the Host-Parasite Interface in <named-content content-type="genus-species">Toxoplasma gondii</named-content>10.1128/mBio.01796-182150-7511https://doaj.org/article/ea7a3291199047328178731b63d09cfa2018-11-01T00:00:00Zhttps://journals.asm.org/doi/10.1128/mBio.01796-18https://doaj.org/toc/2150-7511ABSTRACT Toxoplasma gondii infects approximately 30% of the world’s population, causing disease primarily during pregnancy and in individuals with weakened immune systems. Toxoplasma secretes and exports effector proteins that modulate the host during infection, and several of these proteins are processed by the Golgi-associated aspartyl protease 5 (ASP5). Here, we identify ASP5 substrates by selectively enriching N-terminally derived peptides from wild-type and Δasp5 parasites. We reveal more than 2,000 unique Toxoplasma N-terminal peptides, mapping to both natural N termini and protease cleavage sites. Several of these peptides mapped directly downstream of the characterized ASP5 cleavage site, arginine-arginine-leucine (RRL). We validate candidates as true ASP5 substrates, revealing they are not processed in parasites lacking ASP5 or in wild-type parasites following mutation of the motif from RRL to ARL. All identified ASP5 substrates are dense granule proteins, and interestingly, none appear to be exported, thus differing from the analogous system in related Plasmodium spp. Instead we show that the majority of substrates reside within the parasitophorous vacuole (PV), and its membrane (the PVM), including two kinases and one phosphatase. We show that genetic deletion of WNG2 leads to attenuation in a mouse model, suggesting that this putative kinase is a new virulence factor in Toxoplasma. Collectively, these data constitute the first in-depth analyses of ASP5 substrates and shed new light on the role of ASP5 as a maturase of dense granule proteins during the Toxoplasma lytic cycle. IMPORTANCE Toxoplasma gondii is one of the most successful human parasites. Central to its success is the arsenal of virulence proteins introduced into the infected host cell. Several of these virulence proteins require direct maturation by the aspartyl protease ASP5, and all require ASP5 for translocation into the host cell, yet the true number of ASP5 substrates and complete repertoire of effectors is currently unknown. Here we selectively enrich N-terminally derived peptides using Terminal Amine Isotopic Labeling of Substrates (TAILS) and use quantitative proteomics to reveal novel ASP5 substrates. We identify, using two different enrichment techniques, new ASP5 substrates and their specific cleavage sites. ASP5 substrates include two kinases and one phosphatase that reside at the host-parasite interface, which are important for infection.Michael J. CoffeyLaura F. DagleySimona SeizovaEugene A. KappGiuseppe InfusiniDavid S. RoosJustin A. BoddeyAndrew I. WebbChristopher J. TonkinAmerican Society for MicrobiologyarticleeffectorproteomicsToxoplasmavirulenceMicrobiologyQR1-502ENmBio, Vol 9, Iss 5 (2018)
institution	DOAJ
collection	DOAJ
language	EN
topic	effector proteomics Toxoplasma virulence Microbiology QR1-502
spellingShingle	effector proteomics Toxoplasma virulence Microbiology QR1-502 Michael J. Coffey Laura F. Dagley Simona Seizova Eugene A. Kapp Giuseppe Infusini David S. Roos Justin A. Boddey Andrew I. Webb Christopher J. Tonkin Aspartyl Protease 5 Matures Dense Granule Proteins That Reside at the Host-Parasite Interface in <named-content content-type="genus-species">Toxoplasma gondii</named-content>
description	ABSTRACT Toxoplasma gondii infects approximately 30% of the world’s population, causing disease primarily during pregnancy and in individuals with weakened immune systems. Toxoplasma secretes and exports effector proteins that modulate the host during infection, and several of these proteins are processed by the Golgi-associated aspartyl protease 5 (ASP5). Here, we identify ASP5 substrates by selectively enriching N-terminally derived peptides from wild-type and Δasp5 parasites. We reveal more than 2,000 unique Toxoplasma N-terminal peptides, mapping to both natural N termini and protease cleavage sites. Several of these peptides mapped directly downstream of the characterized ASP5 cleavage site, arginine-arginine-leucine (RRL). We validate candidates as true ASP5 substrates, revealing they are not processed in parasites lacking ASP5 or in wild-type parasites following mutation of the motif from RRL to ARL. All identified ASP5 substrates are dense granule proteins, and interestingly, none appear to be exported, thus differing from the analogous system in related Plasmodium spp. Instead we show that the majority of substrates reside within the parasitophorous vacuole (PV), and its membrane (the PVM), including two kinases and one phosphatase. We show that genetic deletion of WNG2 leads to attenuation in a mouse model, suggesting that this putative kinase is a new virulence factor in Toxoplasma. Collectively, these data constitute the first in-depth analyses of ASP5 substrates and shed new light on the role of ASP5 as a maturase of dense granule proteins during the Toxoplasma lytic cycle. IMPORTANCE Toxoplasma gondii is one of the most successful human parasites. Central to its success is the arsenal of virulence proteins introduced into the infected host cell. Several of these virulence proteins require direct maturation by the aspartyl protease ASP5, and all require ASP5 for translocation into the host cell, yet the true number of ASP5 substrates and complete repertoire of effectors is currently unknown. Here we selectively enrich N-terminally derived peptides using Terminal Amine Isotopic Labeling of Substrates (TAILS) and use quantitative proteomics to reveal novel ASP5 substrates. We identify, using two different enrichment techniques, new ASP5 substrates and their specific cleavage sites. ASP5 substrates include two kinases and one phosphatase that reside at the host-parasite interface, which are important for infection.
format	article
author	Michael J. Coffey Laura F. Dagley Simona Seizova Eugene A. Kapp Giuseppe Infusini David S. Roos Justin A. Boddey Andrew I. Webb Christopher J. Tonkin
author_facet	Michael J. Coffey Laura F. Dagley Simona Seizova Eugene A. Kapp Giuseppe Infusini David S. Roos Justin A. Boddey Andrew I. Webb Christopher J. Tonkin
author_sort	Michael J. Coffey
title	Aspartyl Protease 5 Matures Dense Granule Proteins That Reside at the Host-Parasite Interface in <named-content content-type="genus-species">Toxoplasma gondii</named-content>
title_short	Aspartyl Protease 5 Matures Dense Granule Proteins That Reside at the Host-Parasite Interface in <named-content content-type="genus-species">Toxoplasma gondii</named-content>
title_full	Aspartyl Protease 5 Matures Dense Granule Proteins That Reside at the Host-Parasite Interface in <named-content content-type="genus-species">Toxoplasma gondii</named-content>
title_fullStr	Aspartyl Protease 5 Matures Dense Granule Proteins That Reside at the Host-Parasite Interface in <named-content content-type="genus-species">Toxoplasma gondii</named-content>
title_full_unstemmed	Aspartyl Protease 5 Matures Dense Granule Proteins That Reside at the Host-Parasite Interface in <named-content content-type="genus-species">Toxoplasma gondii</named-content>
title_sort	aspartyl protease 5 matures dense granule proteins that reside at the host-parasite interface in <named-content content-type="genus-species">toxoplasma gondii</named-content>
publisher	American Society for Microbiology
publishDate	2018
url	https://doaj.org/article/ea7a3291199047328178731b63d09cfa
work_keys_str_mv	AT michaeljcoffey aspartylprotease5maturesdensegranuleproteinsthatresideatthehostparasiteinterfaceinnamedcontentcontenttypegenusspeciestoxoplasmagondiinamedcontent AT laurafdagley aspartylprotease5maturesdensegranuleproteinsthatresideatthehostparasiteinterfaceinnamedcontentcontenttypegenusspeciestoxoplasmagondiinamedcontent AT simonaseizova aspartylprotease5maturesdensegranuleproteinsthatresideatthehostparasiteinterfaceinnamedcontentcontenttypegenusspeciestoxoplasmagondiinamedcontent AT eugeneakapp aspartylprotease5maturesdensegranuleproteinsthatresideatthehostparasiteinterfaceinnamedcontentcontenttypegenusspeciestoxoplasmagondiinamedcontent AT giuseppeinfusini aspartylprotease5maturesdensegranuleproteinsthatresideatthehostparasiteinterfaceinnamedcontentcontenttypegenusspeciestoxoplasmagondiinamedcontent AT davidsroos aspartylprotease5maturesdensegranuleproteinsthatresideatthehostparasiteinterfaceinnamedcontentcontenttypegenusspeciestoxoplasmagondiinamedcontent AT justinaboddey aspartylprotease5maturesdensegranuleproteinsthatresideatthehostparasiteinterfaceinnamedcontentcontenttypegenusspeciestoxoplasmagondiinamedcontent AT andrewiwebb aspartylprotease5maturesdensegranuleproteinsthatresideatthehostparasiteinterfaceinnamedcontentcontenttypegenusspeciestoxoplasmagondiinamedcontent AT christopherjtonkin aspartylprotease5maturesdensegranuleproteinsthatresideatthehostparasiteinterfaceinnamedcontentcontenttypegenusspeciestoxoplasmagondiinamedcontent
_version_	1718427062964846592

Aspartyl Protease 5 Matures Dense Granule Proteins That Reside at the Host-Parasite Interface in <named-content content-type="genus-species">Toxoplasma gondii</named-content>

Ejemplares similares