Detection of the Rhoptry Neck Protein Complex in <italic toggle="yes">Plasmodium</italic> Sporozoites and Its Contribution to Sporozoite Invasion of Salivary Glands

ABSTRACT In the Plasmodium life cycle, two infectious stages of parasites, merozoites and sporozoites, share rhoptry and microneme apical structures. A crucial step during merozoite invasion of erythrocytes is the discharge to the host cell membrane of some rhoptry neck proteins as a complex, follow...

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Autores principales:	Mamoru Nozaki, Minami Baba, Mayumi Tachibana, Naohito Tokunaga, Motomi Torii, Tomoko Ishino
Formato:	article
Lenguaje:	EN
Publicado:	American Society for Microbiology 2020
Materias:	sporozoite rhoptry RON complex salivary gland Plasmodium invasion Microbiology QR1-502
Acceso en línea:	https://doaj.org/article/b8938d4de65e40aca9cd91e4fd681164
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spelling	oai:doaj.org-article:b8938d4de65e40aca9cd91e4fd6811642021-11-15T15:30:50ZDetection of the Rhoptry Neck Protein Complex in <italic toggle="yes">Plasmodium</italic> Sporozoites and Its Contribution to Sporozoite Invasion of Salivary Glands10.1128/mSphere.00325-202379-5042https://doaj.org/article/b8938d4de65e40aca9cd91e4fd6811642020-08-01T00:00:00Zhttps://journals.asm.org/doi/10.1128/mSphere.00325-20https://doaj.org/toc/2379-5042ABSTRACT In the Plasmodium life cycle, two infectious stages of parasites, merozoites and sporozoites, share rhoptry and microneme apical structures. A crucial step during merozoite invasion of erythrocytes is the discharge to the host cell membrane of some rhoptry neck proteins as a complex, followed by the formation of a moving junction involving the parasite-secreted protein AMA1 on the parasite membrane. Components of the merozoite rhoptry neck protein complex are also expressed in sporozoites, namely, RON2, RON4, and RON5, suggesting that invasion mechanism elements might be conserved between these infective stages. Recently, we demonstrated that RON2 is required for sporozoite invasion of mosquito salivary gland cells and mammalian hepatocytes, using a sporozoite stage-specific gene knockdown strategy in the rodent malaria parasite model, Plasmodium berghei. Here, we use a coimmunoprecipitation assay and oocyst-derived sporozoite extracts to demonstrate that RON2, RON4, and RON5 also form a complex in sporozoites. The sporozoite stage-specific gene knockdown strategy revealed that both RON4 and RON5 have crucial roles during sporozoite invasion of salivary glands, including a significantly reduced attachment ability required for the onset of gliding. Further analyses indicated that RON2 and RON4 reciprocally affect trafficking to rhoptries in developing sporozoites, while RON5 is independently transported. These findings indicate that the interaction between RON2 and RON4 contributes to their stability and trafficking to rhoptries, in addition to involvement in sporozoite attachment. IMPORTANCE Sporozoites are the motile infectious stage that mediates malaria parasite transmission from mosquitoes to the mammalian host. This study addresses the question whether the rhoptry neck protein complex forms and functions in sporozoites, in addition to its role in merozoites. By applying coimmunoprecipitation and sporozoite stage-specific gene knockdown assays, it was demonstrated that RON2, RON4, and RON5 form a complex and are involved in sporozoite invasion of salivary glands via their attachment ability. These findings shed light on the conserved invasion mechanisms among apicomplexan infective stages. In addition, the sporozoite stage-specific gene knockdown system has revealed for the first time in Plasmodium that the RON2 and RON4 interaction reciprocally affects their stability and trafficking to rhoptries. Our study raises the possibility that the RON complex functions during sporozoite maturation as well as migration toward and invasion of target cells.Mamoru NozakiMinami BabaMayumi TachibanaNaohito TokunagaMotomi ToriiTomoko IshinoAmerican Society for MicrobiologyarticlesporozoiterhoptryRON complexsalivary glandPlasmodiuminvasionMicrobiologyQR1-502ENmSphere, Vol 5, Iss 4 (2020)
institution	DOAJ
collection	DOAJ
language	EN
topic	sporozoite rhoptry RON complex salivary gland Plasmodium invasion Microbiology QR1-502
spellingShingle	sporozoite rhoptry RON complex salivary gland Plasmodium invasion Microbiology QR1-502 Mamoru Nozaki Minami Baba Mayumi Tachibana Naohito Tokunaga Motomi Torii Tomoko Ishino Detection of the Rhoptry Neck Protein Complex in <italic toggle="yes">Plasmodium</italic> Sporozoites and Its Contribution to Sporozoite Invasion of Salivary Glands
description	ABSTRACT In the Plasmodium life cycle, two infectious stages of parasites, merozoites and sporozoites, share rhoptry and microneme apical structures. A crucial step during merozoite invasion of erythrocytes is the discharge to the host cell membrane of some rhoptry neck proteins as a complex, followed by the formation of a moving junction involving the parasite-secreted protein AMA1 on the parasite membrane. Components of the merozoite rhoptry neck protein complex are also expressed in sporozoites, namely, RON2, RON4, and RON5, suggesting that invasion mechanism elements might be conserved between these infective stages. Recently, we demonstrated that RON2 is required for sporozoite invasion of mosquito salivary gland cells and mammalian hepatocytes, using a sporozoite stage-specific gene knockdown strategy in the rodent malaria parasite model, Plasmodium berghei. Here, we use a coimmunoprecipitation assay and oocyst-derived sporozoite extracts to demonstrate that RON2, RON4, and RON5 also form a complex in sporozoites. The sporozoite stage-specific gene knockdown strategy revealed that both RON4 and RON5 have crucial roles during sporozoite invasion of salivary glands, including a significantly reduced attachment ability required for the onset of gliding. Further analyses indicated that RON2 and RON4 reciprocally affect trafficking to rhoptries in developing sporozoites, while RON5 is independently transported. These findings indicate that the interaction between RON2 and RON4 contributes to their stability and trafficking to rhoptries, in addition to involvement in sporozoite attachment. IMPORTANCE Sporozoites are the motile infectious stage that mediates malaria parasite transmission from mosquitoes to the mammalian host. This study addresses the question whether the rhoptry neck protein complex forms and functions in sporozoites, in addition to its role in merozoites. By applying coimmunoprecipitation and sporozoite stage-specific gene knockdown assays, it was demonstrated that RON2, RON4, and RON5 form a complex and are involved in sporozoite invasion of salivary glands via their attachment ability. These findings shed light on the conserved invasion mechanisms among apicomplexan infective stages. In addition, the sporozoite stage-specific gene knockdown system has revealed for the first time in Plasmodium that the RON2 and RON4 interaction reciprocally affects their stability and trafficking to rhoptries. Our study raises the possibility that the RON complex functions during sporozoite maturation as well as migration toward and invasion of target cells.
format	article
author	Mamoru Nozaki Minami Baba Mayumi Tachibana Naohito Tokunaga Motomi Torii Tomoko Ishino
author_facet	Mamoru Nozaki Minami Baba Mayumi Tachibana Naohito Tokunaga Motomi Torii Tomoko Ishino
author_sort	Mamoru Nozaki
title	Detection of the Rhoptry Neck Protein Complex in <italic toggle="yes">Plasmodium</italic> Sporozoites and Its Contribution to Sporozoite Invasion of Salivary Glands
title_short	Detection of the Rhoptry Neck Protein Complex in <italic toggle="yes">Plasmodium</italic> Sporozoites and Its Contribution to Sporozoite Invasion of Salivary Glands
title_full	Detection of the Rhoptry Neck Protein Complex in <italic toggle="yes">Plasmodium</italic> Sporozoites and Its Contribution to Sporozoite Invasion of Salivary Glands
title_fullStr	Detection of the Rhoptry Neck Protein Complex in <italic toggle="yes">Plasmodium</italic> Sporozoites and Its Contribution to Sporozoite Invasion of Salivary Glands
title_full_unstemmed	Detection of the Rhoptry Neck Protein Complex in <italic toggle="yes">Plasmodium</italic> Sporozoites and Its Contribution to Sporozoite Invasion of Salivary Glands
title_sort	detection of the rhoptry neck protein complex in <italic toggle="yes">plasmodium</italic> sporozoites and its contribution to sporozoite invasion of salivary glands
publisher	American Society for Microbiology
publishDate	2020
url	https://doaj.org/article/b8938d4de65e40aca9cd91e4fd681164
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Detection of the Rhoptry Neck Protein Complex in <italic toggle="yes">Plasmodium</italic> Sporozoites and Its Contribution to Sporozoite Invasion of Salivary Glands

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