Simultaneous Ribosome Profiling of Human Host Cells Infected with <named-content content-type="genus-species">Toxoplasma gondii</named-content>

ABSTRACT Toxoplasma gondii is a ubiquitous obligate intracellular parasite that infects the nucleated cells of warm-blooded animals. From within the parasitophorous vacuole in which they reside, Toxoplasma tachyzoites secrete an arsenal of effector proteins that can reprogram host gene expression to...

Descripción completa

Guardado en:

Detalles Bibliográficos
Autores principales:	Michael J. Holmes, Premal Shah, Ronald C. Wek, William J. Sullivan
Formato:	article
Lenguaje:	EN
Publicado:	American Society for Microbiology 2019
Materias:	ribosomal profiling Toxoplasma apicomplexa parasites translation translation control Microbiology QR1-502
Acceso en línea:	https://doaj.org/article/66647780fabc4a5db5d2da1e3f7b81f0
Etiquetas:	Agregar Etiqueta Sin Etiquetas, Sea el primero en etiquetar este registro!

id	oai:doaj.org-article:66647780fabc4a5db5d2da1e3f7b81f0
record_format	dspace
spelling	oai:doaj.org-article:66647780fabc4a5db5d2da1e3f7b81f02021-11-15T15:22:20ZSimultaneous Ribosome Profiling of Human Host Cells Infected with <named-content content-type="genus-species">Toxoplasma gondii</named-content>10.1128/mSphere.00292-192379-5042https://doaj.org/article/66647780fabc4a5db5d2da1e3f7b81f02019-06-01T00:00:00Zhttps://journals.asm.org/doi/10.1128/mSphere.00292-19https://doaj.org/toc/2379-5042ABSTRACT Toxoplasma gondii is a ubiquitous obligate intracellular parasite that infects the nucleated cells of warm-blooded animals. From within the parasitophorous vacuole in which they reside, Toxoplasma tachyzoites secrete an arsenal of effector proteins that can reprogram host gene expression to facilitate parasite survival and replication. Gaining a better understanding of how host gene expression is altered upon infection is central for understanding parasite strategies for host invasion and for developing new parasite therapies. Here, we applied ribosome profiling coupled with mRNA measurements to concurrently study gene expression in the parasite and in host human foreskin fibroblasts. By examining the parasite transcriptome and translatome, we identified potential upstream open reading frames that may permit the stress-induced preferential translation of parasite mRNAs. We also determined that tachyzoites reduce host death-associated pathways and increase survival, proliferation, and motility in both quiescent and proliferative host cell models of infection. Additionally, proliferative cells alter their gene expression in ways that are consistent with massive transcriptional rewiring, while quiescent cells were best characterized by reentry into the cell cycle. We also identified a translational control regimen consistent with mechanistic target of rapamycin (mTOR) activation in quiescent cells and, to a lesser degree, in proliferative cells. This study illustrates the utility of the method for dissection of gene expression programs simultaneously in the parasite and host. IMPORTANCE Toxoplasma gondii is a single-celled parasite that has infected up to one-third of the world’s population. Significant overhauls in gene expression in both the parasite and the host cell accompany parasite invasion, and a better understanding of these changes may lead to the development of new therapeutic agents. In this study, we employed ribosome profiling to determine the changes that occur at the levels of transcription and translation in both the parasite and the infected host cell at the same time. We discovered features of Toxoplasma mRNAs that suggest a means for controlling parasite gene expression under stressful conditions. We also show that differences in host gene expression occur depending on whether they are confluent or not. Our findings demonstrate the feasibility of using ribosomal profiling to interrogate the host-parasite dynamic under a variety of conditions.Michael J. HolmesPremal ShahRonald C. WekWilliam J. SullivanAmerican Society for Microbiologyarticleribosomal profilingToxoplasmaapicomplexaparasitestranslationtranslation controlMicrobiologyQR1-502ENmSphere, Vol 4, Iss 3 (2019)
institution	DOAJ
collection	DOAJ
language	EN
topic	ribosomal profiling Toxoplasma apicomplexa parasites translation translation control Microbiology QR1-502
spellingShingle	ribosomal profiling Toxoplasma apicomplexa parasites translation translation control Microbiology QR1-502 Michael J. Holmes Premal Shah Ronald C. Wek William J. Sullivan Simultaneous Ribosome Profiling of Human Host Cells Infected with <named-content content-type="genus-species">Toxoplasma gondii</named-content>
description	ABSTRACT Toxoplasma gondii is a ubiquitous obligate intracellular parasite that infects the nucleated cells of warm-blooded animals. From within the parasitophorous vacuole in which they reside, Toxoplasma tachyzoites secrete an arsenal of effector proteins that can reprogram host gene expression to facilitate parasite survival and replication. Gaining a better understanding of how host gene expression is altered upon infection is central for understanding parasite strategies for host invasion and for developing new parasite therapies. Here, we applied ribosome profiling coupled with mRNA measurements to concurrently study gene expression in the parasite and in host human foreskin fibroblasts. By examining the parasite transcriptome and translatome, we identified potential upstream open reading frames that may permit the stress-induced preferential translation of parasite mRNAs. We also determined that tachyzoites reduce host death-associated pathways and increase survival, proliferation, and motility in both quiescent and proliferative host cell models of infection. Additionally, proliferative cells alter their gene expression in ways that are consistent with massive transcriptional rewiring, while quiescent cells were best characterized by reentry into the cell cycle. We also identified a translational control regimen consistent with mechanistic target of rapamycin (mTOR) activation in quiescent cells and, to a lesser degree, in proliferative cells. This study illustrates the utility of the method for dissection of gene expression programs simultaneously in the parasite and host. IMPORTANCE Toxoplasma gondii is a single-celled parasite that has infected up to one-third of the world’s population. Significant overhauls in gene expression in both the parasite and the host cell accompany parasite invasion, and a better understanding of these changes may lead to the development of new therapeutic agents. In this study, we employed ribosome profiling to determine the changes that occur at the levels of transcription and translation in both the parasite and the infected host cell at the same time. We discovered features of Toxoplasma mRNAs that suggest a means for controlling parasite gene expression under stressful conditions. We also show that differences in host gene expression occur depending on whether they are confluent or not. Our findings demonstrate the feasibility of using ribosomal profiling to interrogate the host-parasite dynamic under a variety of conditions.
format	article
author	Michael J. Holmes Premal Shah Ronald C. Wek William J. Sullivan
author_facet	Michael J. Holmes Premal Shah Ronald C. Wek William J. Sullivan
author_sort	Michael J. Holmes
title	Simultaneous Ribosome Profiling of Human Host Cells Infected with <named-content content-type="genus-species">Toxoplasma gondii</named-content>
title_short	Simultaneous Ribosome Profiling of Human Host Cells Infected with <named-content content-type="genus-species">Toxoplasma gondii</named-content>
title_full	Simultaneous Ribosome Profiling of Human Host Cells Infected with <named-content content-type="genus-species">Toxoplasma gondii</named-content>
title_fullStr	Simultaneous Ribosome Profiling of Human Host Cells Infected with <named-content content-type="genus-species">Toxoplasma gondii</named-content>
title_full_unstemmed	Simultaneous Ribosome Profiling of Human Host Cells Infected with <named-content content-type="genus-species">Toxoplasma gondii</named-content>
title_sort	simultaneous ribosome profiling of human host cells infected with <named-content content-type="genus-species">toxoplasma gondii</named-content>
publisher	American Society for Microbiology
publishDate	2019
url	https://doaj.org/article/66647780fabc4a5db5d2da1e3f7b81f0
work_keys_str_mv	AT michaeljholmes simultaneousribosomeprofilingofhumanhostcellsinfectedwithnamedcontentcontenttypegenusspeciestoxoplasmagondiinamedcontent AT premalshah simultaneousribosomeprofilingofhumanhostcellsinfectedwithnamedcontentcontenttypegenusspeciestoxoplasmagondiinamedcontent AT ronaldcwek simultaneousribosomeprofilingofhumanhostcellsinfectedwithnamedcontentcontenttypegenusspeciestoxoplasmagondiinamedcontent AT williamjsullivan simultaneousribosomeprofilingofhumanhostcellsinfectedwithnamedcontentcontenttypegenusspeciestoxoplasmagondiinamedcontent
_version_	1718428011531862016

Simultaneous Ribosome Profiling of Human Host Cells Infected with <named-content content-type="genus-species">Toxoplasma gondii</named-content>

Ejemplares similares