{"title":"GRA47 对弓形虫寄生泡的形态和渗透性非常重要。","authors":"","doi":"10.1016/j.ijpara.2024.06.003","DOIUrl":null,"url":null,"abstract":"<div><p>Establishing an intact intracellular parasitophorous vacuole (PV) that enables efficient nutrient uptake and protein trafficking is essential for the survival and proliferation of <span><em>Toxoplasma gondii</em></span><span>. Although the PV membrane (PVM)-localized dense granule protein 17 (GRA17) and GRA23 mediate the permeability of the PVM to small molecules, including nutrient uptake and excretion of metabolic by-products, the molecular mechanism by which </span><em>T. gondii</em> acquires nutrients remains unclear. In this study, we showed that the secreted protein GRA47 contributed to normal PV morphology, PVM permeability to small molecules, growth, and virulence in <em>T. gondii</em>. Co-immunoprecipitation analysis demonstrated potential interaction of GRA47 with GRA72, and the loss of GRA72 affected PV morphology, parasite growth and infectivity. To investigate the biological relationship among GRA47, GRA72, GRA17 and GRA23, attempts were made to construct strains with double gene deletion and overexpressing strains. Only Δ<em>gra</em>23Δ<em>gra72</em> was successfully constructed. This strain exhibited a significant increase in the proportion of aberrant PVs compared with the Δ<em>gra23</em> strain. Overexpressing one of the three related GRAs partially rescued PVs with aberrant morphology in Δ<em>gra47</em>, Δ<em>gra72</em> and Δ<em>gra17</em>, while the expression of the <em>Plasmodium falciparum</em> PVM protein <em>Pf</em>Exp2, an ortholog of GRA17 and GRA23, fully rescued the PV morphological defect in all three Δ<em>gra</em> strains. These results suggest that these GRA proteins may not be functionally redundant but rather work in different ways to regulate nutrient acquisition. These findings highlight the versatility of the nutrient uptake mechanisms in <em>T. gondii</em>, which may contribute to the parasite’s remarkable ability to grow in different cellular niches in a very broad range of hosts.</p></div>","PeriodicalId":13725,"journal":{"name":"International journal for parasitology","volume":null,"pages":null},"PeriodicalIF":3.7000,"publicationDate":"2024-09-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":"0","resultStr":"{\"title\":\"GRA47 is important for the morphology and permeability of the parasitophorous vacuole in Toxoplasma gondii\",\"authors\":\"\",\"doi\":\"10.1016/j.ijpara.2024.06.003\",\"DOIUrl\":null,\"url\":null,\"abstract\":\"<div><p>Establishing an intact intracellular parasitophorous vacuole (PV) that enables efficient nutrient uptake and protein trafficking is essential for the survival and proliferation of <span><em>Toxoplasma gondii</em></span><span>. Although the PV membrane (PVM)-localized dense granule protein 17 (GRA17) and GRA23 mediate the permeability of the PVM to small molecules, including nutrient uptake and excretion of metabolic by-products, the molecular mechanism by which </span><em>T. gondii</em> acquires nutrients remains unclear. In this study, we showed that the secreted protein GRA47 contributed to normal PV morphology, PVM permeability to small molecules, growth, and virulence in <em>T. gondii</em>. Co-immunoprecipitation analysis demonstrated potential interaction of GRA47 with GRA72, and the loss of GRA72 affected PV morphology, parasite growth and infectivity. To investigate the biological relationship among GRA47, GRA72, GRA17 and GRA23, attempts were made to construct strains with double gene deletion and overexpressing strains. Only Δ<em>gra</em>23Δ<em>gra72</em> was successfully constructed. This strain exhibited a significant increase in the proportion of aberrant PVs compared with the Δ<em>gra23</em> strain. Overexpressing one of the three related GRAs partially rescued PVs with aberrant morphology in Δ<em>gra47</em>, Δ<em>gra72</em> and Δ<em>gra17</em>, while the expression of the <em>Plasmodium falciparum</em> PVM protein <em>Pf</em>Exp2, an ortholog of GRA17 and GRA23, fully rescued the PV morphological defect in all three Δ<em>gra</em> strains. These results suggest that these GRA proteins may not be functionally redundant but rather work in different ways to regulate nutrient acquisition. These findings highlight the versatility of the nutrient uptake mechanisms in <em>T. gondii</em>, which may contribute to the parasite’s remarkable ability to grow in different cellular niches in a very broad range of hosts.</p></div>\",\"PeriodicalId\":13725,\"journal\":{\"name\":\"International journal for parasitology\",\"volume\":null,\"pages\":null},\"PeriodicalIF\":3.7000,\"publicationDate\":\"2024-09-01\",\"publicationTypes\":\"Journal Article\",\"fieldsOfStudy\":null,\"isOpenAccess\":false,\"openAccessPdf\":\"\",\"citationCount\":\"0\",\"resultStr\":null,\"platform\":\"Semanticscholar\",\"paperid\":null,\"PeriodicalName\":\"International journal for parasitology\",\"FirstCategoryId\":\"3\",\"ListUrlMain\":\"https://www.sciencedirect.com/science/article/pii/S0020751924001358\",\"RegionNum\":2,\"RegionCategory\":\"医学\",\"ArticlePicture\":[],\"TitleCN\":null,\"AbstractTextCN\":null,\"PMCID\":null,\"EPubDate\":\"\",\"PubModel\":\"\",\"JCR\":\"Q1\",\"JCRName\":\"PARASITOLOGY\",\"Score\":null,\"Total\":0}","platform":"Semanticscholar","paperid":null,"PeriodicalName":"International journal for parasitology","FirstCategoryId":"3","ListUrlMain":"https://www.sciencedirect.com/science/article/pii/S0020751924001358","RegionNum":2,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":null,"EPubDate":"","PubModel":"","JCR":"Q1","JCRName":"PARASITOLOGY","Score":null,"Total":0}
GRA47 is important for the morphology and permeability of the parasitophorous vacuole in Toxoplasma gondii
Establishing an intact intracellular parasitophorous vacuole (PV) that enables efficient nutrient uptake and protein trafficking is essential for the survival and proliferation of Toxoplasma gondii. Although the PV membrane (PVM)-localized dense granule protein 17 (GRA17) and GRA23 mediate the permeability of the PVM to small molecules, including nutrient uptake and excretion of metabolic by-products, the molecular mechanism by which T. gondii acquires nutrients remains unclear. In this study, we showed that the secreted protein GRA47 contributed to normal PV morphology, PVM permeability to small molecules, growth, and virulence in T. gondii. Co-immunoprecipitation analysis demonstrated potential interaction of GRA47 with GRA72, and the loss of GRA72 affected PV morphology, parasite growth and infectivity. To investigate the biological relationship among GRA47, GRA72, GRA17 and GRA23, attempts were made to construct strains with double gene deletion and overexpressing strains. Only Δgra23Δgra72 was successfully constructed. This strain exhibited a significant increase in the proportion of aberrant PVs compared with the Δgra23 strain. Overexpressing one of the three related GRAs partially rescued PVs with aberrant morphology in Δgra47, Δgra72 and Δgra17, while the expression of the Plasmodium falciparum PVM protein PfExp2, an ortholog of GRA17 and GRA23, fully rescued the PV morphological defect in all three Δgra strains. These results suggest that these GRA proteins may not be functionally redundant but rather work in different ways to regulate nutrient acquisition. These findings highlight the versatility of the nutrient uptake mechanisms in T. gondii, which may contribute to the parasite’s remarkable ability to grow in different cellular niches in a very broad range of hosts.
期刊介绍:
International Journal for Parasitology offers authors the option to sponsor nonsubscriber access to their articles on Elsevier electronic publishing platforms. For more information please view our Sponsored Articles page. The International Journal for Parasitology publishes the results of original research in all aspects of basic and applied parasitology, including all the fields covered by its Specialist Editors, and ranging from parasites and host-parasite relationships of intrinsic biological interest to those of social and economic importance in human and veterinary medicine and agriculture.