A monoclonal antibody against Toxoplasma gondii of Tg556 clone (Tg556) blotted a 29 kDa protein, which was localized in the dense granules of tachyzoites and secreted into the parasitophorous vacuolar membrane (PVM) after infection to host cells. A cDNA fragment encoding the protein was obtained by screening a T. gondii cDNA expression library with Tg556, and the full-length was completed by 5'-RACE of 2,086 bp containing an open reading frame (ORF) of 669 bp. The ORF encoded a polypeptide of 222 amino acids homologous to the revised GRA3 but not to the first reported one. The polypeptide has 3 hydrophobic moieties of an N-terminal stop transfer sequence and 2 transmembrane domains (TMD) in posterior half of the sequence, a cytoplasmic localization motif after the second TMD and an endoplasmic reticulum (ER) retrival motif in the C-terminal end, which suggests GRA3 as a type III transmembrane protein. With the ORF of GRA3, yeast two-hybrid assay was performed in HeLa cDNA expression library, which resulted in the interaction of GRA3 with calcium modulating ligand (CAMLG), a type II transmembrane protein of ER. The specific binding of GRA3 and CAMLG was confirmed by glutathione S-transferase (GST) pull-down and immunoprecipitation assays. The localities of fluorescence transfectionally expressed from GRA3 and CAMLG plasmids were overlapped completely in HeLa cell cytoplasm. In immunofluorescence assay, GRA3 and CAMLG were shown to be co-localized in the PVM of host cells. Structural binding of PVM-inserted GRA3 to CAMLG of ER suggested the receptor-ligand of ER recruitment to PVM during the parasitism of T. gondii.
Citations
Citations to this article as recorded by
Toxoplasma effector TgROP1 establishes membrane contact sites with the endoplasmic reticulum during infection Chahat Mehra, Jesús Alvarado Valverde, Ana Margarida Nogueira Matias, Francesca Torelli, Tânia Catarina Medeiros, Julian Straub, James D. Asaki, Peter J. Bradley, Katja Luck, Steffen Lawo, Moritz Treeck, Lena Pernas Nature Microbiology.2025; 10(12): 3331. CrossRef
Toxoplasma–host endoplasmic reticulum interaction: How T. gondii activates unfolded protein response and modulates immune response Obed Cudjoe, Roger Afful, Tonny Abraham Hagan Current Research in Microbial Sciences.2024; 6: 100223. CrossRef
Host cell manipulation by microsporidia secreted effectors: Insights into intracellular pathogenesis Liyuan Tang, Musa Makongoro Sabi, Ming Fu, Jingyu Guan, Yongliang Wang, Tian Xia, Kai Zheng, Hongnan Qu, Bing Han Journal of Eukaryotic Microbiology.2024;[Epub] CrossRef
Preparation and Preliminary Application of Epitope Peptide-Based Antibody against Toxoplasma gondii GRA3 Ru Wang, Minmin Wu, Haijian Cai, Ran An, Ying Chen, Jie Wang, Nan Zhou, Jian Du Tropical Medicine and Infectious Disease.2023; 8(3): 143. CrossRef
Intracellular life of protozoan Toxoplasma gondii: Parasitophorous vacuole establishment and survival strategies JULIANA A. PORTES, ROSSIANE C. VOMMARO, LUCIO AYRES CALDAS, ERICA S. MARTINS-DUARTE BIOCELL.2023; 47(4): 929. CrossRef
A state-of-the-art methodology for high-throughput in silico vaccine discovery against protozoan parasites and exemplified with discovered candidates for Toxoplasma gondii Stephen J. Goodswen, Paul J. Kennedy, John T. Ellis Scientific Reports.2023;[Epub] CrossRef
Toxoplasma gondii dense granule protein 3 promotes endoplasmic reticulum stress-induced apoptosis by activating the PERK pathway Cudjoe Obed, Minmin Wu, Ying Chen, Ran An, Haijian Cai, Qingli Luo, Li Yu, Jie Wang, Fang Liu, Jilong Shen, Jian Du Parasites & Vectors.2022;[Epub] CrossRef
The UPR sensor IRE1α promotes dendritic cell responses to control Toxoplasma gondii infection Anaïs F Poncet, Victor Bosteels, Eik Hoffmann, Sylia Chehade, Sofie Rennen, Ludovic Huot, Véronique Peucelle, Sandra Maréchal, Jamal Khalife, Nicolas Blanchard, Sophie Janssens, Sabrina Marion EMBO reports.2021;[Epub] CrossRef
Interorganellar communication and membrane contact sites in protozoan parasites Herbert J. Santos, Tomoyoshi Nozaki Parasitology International.2021; 83: 102372. CrossRef
A Newly Discovered Dense Granule Protein 3 in Neospora caninum Panpan Zhao, Jingquan Dong, Lili Cao, Jianhua Li, Xichen Zhang, Xin Li, Xiaocen Wang, Babatunde Kazeem Bello, Nan Zhang, Pengtao Gong Acta Parasitologica.2021; 66(4): 1581. CrossRef
Manipulation of Host Cell Organelles by Intracellular Pathogens Malte Kellermann, Felix Scharte, Michael Hensel International Journal of Molecular Sciences.2021; 22(12): 6484. CrossRef
Toxoplasma gondii Parasitophorous Vacuole Membrane-Associated Dense Granule Proteins Regulate Maturation of the Cyst Wall Rebekah B. Guevara, Barbara A. Fox, David J. Bzik, Silvia N. J. Moreno mSphere.2020;[Epub] CrossRef
TheToxoplasma gondiidense granule protein TgGRA3 interacts with host Golgi and dysregulates anterograde transport Maika S. Deffieu, Tchilabalo Dilezitoko Alayi, Christian Slomianny, Stanislas Tomavo Biology Open.2019;[Epub] CrossRef
Rhoptry and Dense Granule Secreted Effectors Regulate CD8+ T Cell Recognition of Toxoplasma gondii Infected Host Cells Leah M. Rommereim, Barbara A. Fox, Kiah L. Butler, Viviana Cantillana, Gregory A. Taylor, David J. Bzik Frontiers in Immunology.2019;[Epub] CrossRef
Identification of two novel host proteins interacting with Toxoplasma gondii 14-3-3 protein by yeast two-hybrid system Fa-Cai Li, Qing Liu, Hany M. Elsheikha, Wen-Bin Yang, Jun-Ling Hou, Xing-Quan Zhu Parasitology Research.2018; 117(4): 1291. CrossRef
The ER phagosome connection in the era of membrane contact sites Paula Nunes-Hasler, Nicolas Demaurex Biochimica et Biophysica Acta (BBA) - Molecular Cell Research.2017; 1864(9): 1513. CrossRef
How Toxoplasma and malaria parasites defy first, then exploit host autophagic and endocytic pathways for growth Isabelle Coppens Current Opinion in Microbiology.2017; 40: 32. CrossRef
Genotyping of polymorphic effectors of Toxoplasma gondii isolates from China Weisheng Cheng, Cong Wang, Ting Xu, Fang Liu, Faustina Pappoe, Qingli Luo, Yuanhong Xu, Fangli Lu, Jilong Shen Parasites & Vectors.2017;[Epub] CrossRef
Apicomplexans pulling the strings: manipulation of the host cell cytoskeleton dynamics RITA CARDOSO, HELENA SOARES, ANDREW HEMPHILL, ALEXANDRE LEITÃO Parasitology.2016; 143(8): 957. CrossRef
Variation detection based on next-generation sequencing of type Chinese 1 strains of Toxoplasma gondii with different virulence from China Weisheng Cheng, Fang Liu, Man Li, Xiaodong Hu, He Chen, Faustina Pappoe, Qingli Luo, Huiqin Wen, Tian Xing, Yuanhong Xu, Jilong Shen BMC Genomics.2015;[Epub] CrossRef
Toxoplasmosis and Epilepsy — Systematic Review and Meta Analysis Edgard B. Ngoungou, Devender Bhalla, Amandine Nzoghe, Marie-Laure Dardé, Pierre-Marie Preux, Rima McLeod PLOS Neglected Tropical Diseases.2015; 9(2): e0003525. CrossRef
Neospora caninum Recruits Host Cell Structures to Its Parasitophorous Vacuole and Salvages Lipids from Organelles Sabrina J. Nolan, Julia D. Romano, Thomas Luechtefeld, Isabelle Coppens Eukaryotic Cell.2015; 14(5): 454. CrossRef
Phylogeny and virulence divergency analyses of Toxoplasma gondii isolates from China Min Li, Xu-Wei Mo, Lin Wang, He Chen, Qing-Li Luo, Hui-Qin Wen, Wei Wei, Ai-Mei Zhang, Jian Du, Fang-Li Lu, Zhao-Rong Lun, Ji-Long Shen Parasites & Vectors.2014;[Epub] CrossRef
High Expression of Water-Soluble Recombinant Antigenic Domains ofToxoplasma gondii Secretory Organelles Zhaoshou Yang, Hye-Jin Ahn, Ho-Woo Nam The Korean Journal of Parasitology.2014; 52(4): 367. CrossRef
Exploitation of auxotrophies and metabolic defects in Toxoplasma as therapeutic approaches Isabelle Coppens International Journal for Parasitology.2014; 44(2): 109. CrossRef
Selective and strain-specific NFAT4 activation by the Toxoplasma gondii polymorphic dense granule protein GRA6 Ji Su Ma, Miwa Sasai, Jun Ohshima, Youngae Lee, Hironori Bando, Kiyoshi Takeda, Masahiro Yamamoto Journal of Experimental Medicine.2014; 211(10): 2013. CrossRef
Discovering a vaccine against neosporosis using computers: is it feasible? Stephen J. Goodswen, Paul J. Kennedy, John T. Ellis Trends in Parasitology.2014; 30(8): 401. CrossRef
Host Organelle Hijackers: a similarmodus operandiforToxoplasma gondiiandChlamydia trachomatis: co-infection model as a tool to investigate pathogenesis Julia D. Romano, Isabelle Coppens Pathogens and Disease.2013; 69(2): 72. CrossRef
Interactions of pathogen-containing compartments with the secretory pathway Johnathan Canton, Peter E. Kima Cellular Microbiology.2012; 14(11): 1676. CrossRef
Transcriptomic Analysis of Toxoplasma Development Reveals Many Novel Functions and Structures Specific to Sporozoites and Oocysts Heather M. Fritz, Kerry R. Buchholz, Xiucui Chen, Blythe Durbin-Johnson, David M. Rocke, Patricia A. Conrad, John C. Boothroyd, Silvia N. Moreno PLoS ONE.2012; 7(2): e29998. CrossRef
Toxoplasma gondii Actively Inhibits Neuronal Function in Chronically Infected Mice Fahad Haroon, Ulrike Händel, Frank Angenstein, Jürgen Goldschmidt, Peter Kreutzmann, Holger Lison, Klaus-Dieter Fischer, Henning Scheich, Wolfram Wetzel, Dirk Schlüter, Eike Budinger, Silvia N. Moreno PLoS ONE.2012; 7(4): e35516. CrossRef
Toxoplasma gondii: Expression of GRA1 gene in endoplasmic reticulum promotes both growth and adherence and modulates intracellular calcium release in macrophages Jie Lin, Xu Lin, Guo-hua Yang, Yong Wang, Bi-wen Peng, Jian-yin Lin Experimental Parasitology.2010; 125(2): 165. CrossRef
Transmembrane and Ubiquitin-Like Domain Containing 1 (Tmub1) Regulates Locomotor Activity and Wakefulness in Mice and Interacts with CAMLG Wandong Zhang, Katerina V. Savelieva, Adisak Suwanichkul, Daniel L. Small, Laura L. Kirkpatrick, Nianhua Xu, Thomas H. Lanthorn, Gui-Lan Ye, Georges Chapouthier PLoS ONE.2010; 5(6): e11261. CrossRef
GRA Proteins of Toxoplasma gondii: Maintenance of Host-Parasite Interactions across the Parasitophorous Vacuolar Membrane Ho-Woo Nam The Korean Journal of Parasitology.2009; 47(Suppl): S29. CrossRef
Interactions between GRA proteins of dense granules in Toxoplasma gondii and host cell proteins were analyzed by yeast two-hybrid technique. The cMyc-GRA fusion proteins expressed from pGBKT7 plasmid in Y187 yeast were bound to host cell proteins from pGADT7-Rec-HeLa cDNA library transformed to AH109 yeast by mating method. By the selection procedures, a total of 939 colonies of the SD/-AHLT culture, 348 colonies of the X-α-gal positive and PCR, 157 colonies of the X-β-gal assay were chosen for sequencing the cDNA and finally 90 colonies containing ORF were selected to analyze the interactions. GRA proteins interacted with a variety of host cell proteins such as enzymes, structural and functional proteins of organellar proteins of broad spectrum. Several specific bindings of each GRA protein to host proteins were discussed presumptively the role of GRA proteins after secreting into the parasitophorous vacuoles (PV) and the PV membrane in the parasitism of this parasite.
Citations
Citations to this article as recorded by
Toxoplasma–host endoplasmic reticulum interaction: How T. gondii activates unfolded protein response and modulates immune response Obed Cudjoe, Roger Afful, Tonny Abraham Hagan Current Research in Microbial Sciences.2024; 6: 100223. CrossRef
Preparation and Preliminary Application of Epitope Peptide-Based Antibody against Toxoplasma gondii GRA3 Ru Wang, Minmin Wu, Haijian Cai, Ran An, Ying Chen, Jie Wang, Nan Zhou, Jian Du Tropical Medicine and Infectious Disease.2023; 8(3): 143. CrossRef
Disruption of Dense Granular Protein 2 (GRA2) Decreases the Virulence of Neospora caninum Jingquan Dong, Nan Zhang, Panpan Zhao, Jianhua Li, Lili Cao, Xiaocen Wang, Xin Li, Ju Yang, Xichen Zhang, Pengtao Gong Frontiers in Veterinary Science.2021;[Epub] CrossRef
Interorganellar communication and membrane contact sites in protozoan parasites Herbert J. Santos, Tomoyoshi Nozaki Parasitology International.2021; 83: 102372. CrossRef
Descriptive Comparison of ELISAs for the Detection of Toxoplasma gondii Antibodies in Animals: A Systematic Review K. L. D. Tharaka D. Liyanage, Anke Wiethoelter, Jasmin Hufschmid, Abdul Jabbar Pathogens.2021; 10(5): 605. CrossRef
Toxoplasma gondii GRA9 Regulates the Activation of NLRP3 Inflammasome to Exert Anti-Septic Effects in Mice Jae-Sung Kim, Seok-Jun Mun, Euni Cho, Donggyu Kim, Wooic Son, Hye-In Jeon, Hyo Keun Kim, Kiseok Jang, Chul-Su Yang International Journal of Molecular Sciences.2020; 21(22): 8437. CrossRef
Identification of two novel host proteins interacting with Toxoplasma gondii 14-3-3 protein by yeast two-hybrid system Fa-Cai Li, Qing Liu, Hany M. Elsheikha, Wen-Bin Yang, Jun-Ling Hou, Xing-Quan Zhu Parasitology Research.2018; 117(4): 1291. CrossRef
Toxoplasma gondii GRA8 induces ATP5A1–SIRT3-mediated mitochondrial metabolic resuscitation: a potential therapy for sepsis Ye-Ram Kim, Jae-Sung Kim, Jin-Seung Yun, Sojin Kim, Sun Young Kim, Kiseok Jang, Chul-Su Yang Experimental & Molecular Medicine.2018; 50(3): e464. CrossRef
Secretory organelle trafficking in Toxoplasma gondii: A long story for a short travel Kannan Venugopal, Sabrina Marion International Journal of Medical Microbiology.2018; 308(7): 751. CrossRef
Measurement of binding strength between prey proteins interacting with Toxoplasma gondii SAG1 and SAG2 using isothermal titration calorimetry (ITC) Meng-Yee Lai, Yee-Ling Lau Acta Parasitologica.2018; 63(1): 106. CrossRef
Toxoplasma gondii GRA7-Targeted ASC and PLD1 Promote Antibacterial Host Defense via PKCα Hyun-Jung Koh, Ye-Ram Kim, Jae-Sung Kim, Jin-Seung Yun, Kiseok Jang, Chul-Su Yang, Sarah M. Fortune PLOS Pathogens.2017; 13(1): e1006126. CrossRef
The Toxoplasma Parasitophorous Vacuole: An Evolving Host–Parasite Frontier Barbara Clough, Eva-Maria Frickel Trends in Parasitology.2017; 33(6): 473. CrossRef
Perturbed human sub-networks by Fusobacterium nucleatum candidate virulence proteins Andreas Zanzoni, Lionel Spinelli, Shérazade Braham, Christine Brun Microbiome.2017;[Epub] CrossRef
Screening and identification of host proteins interacting with Toxoplasma gondii SAG2 by yeast two-hybrid assay Meng-Yee Lai, Yee-Ling Lau Parasites & Vectors.2017;[Epub] CrossRef
Transcriptional changes of mouse splenocyte organelle components following acute infection with Toxoplasma gondii Jun-Jun He, Jun Ma, Fa-Cai Li, Hui-Qun Song, Min-Jun Xu, Xing-Quan Zhu Experimental Parasitology.2016; 167: 7. CrossRef
Toxoplasma gondii GRA7-Induced TRAF6 Activation Contributes to Host Protective Immunity Chul-Su Yang, Jae-Min Yuk, Young-Ha Lee, Eun-Kyeong Jo, J. H. Adams Infection and Immunity.2016; 84(1): 339. CrossRef
Evaluation of an indirect ELISA using recombinant granule antigen GRA1, GRA7 and soluble antigens for serodiagnosis of Toxoplasma gondii infection in chickens Xuefei Sun, Zedong Wang, Jiping Li, Feng Wei, Quan Liu Research in Veterinary Science.2015; 100: 161. CrossRef
Comparative mapping of host–pathogen protein–protein interactions Priya S Shah, Jason A Wojcechowskyj, Manon Eckhardt, Nevan J Krogan Current Opinion in Microbiology.2015; 27: 62. CrossRef
Selective and strain-specific NFAT4 activation by the Toxoplasma gondii polymorphic dense granule protein GRA6 Ji Su Ma, Miwa Sasai, Jun Ohshima, Youngae Lee, Hironori Bando, Kiyoshi Takeda, Masahiro Yamamoto Journal of Experimental Medicine.2014; 211(10): 2013. CrossRef
Recombinant Dense Granular Protein (GRA5) for Detection of Human Toxoplasmosis by Western Blot Xiao Teng Ching, Yee Ling Lau, Mun Yik Fong, Veeranoot Nissapatorn, Hemah Andiappan BioMed Research International.2014; 2014: 1. CrossRef
A novel dense granule protein, GRA22, is involved in regulating parasite egress in Toxoplasma gondii Tadashi Okada, Dini Marmansari, Zeng-mei Li, Altanchimeg Adilbish, Shishenkov Canko, Akio Ueno, Haruhi Shono, Hidefumi Furuoka, Makoto Igarashi Molecular and Biochemical Parasitology.2013; 189(1-2): 5. CrossRef
GRA 14, a novel dense granule protein from Neospora caninum Gongzhen Liu, Xia Cui, Pan Hao, Daoyu Yang, Jing Liu, Qun Liu Acta Biochimica et Biophysica Sinica.2013; 45(7): 607. CrossRef
Transcriptomic Analysis of Toxoplasma Development Reveals Many Novel Functions and Structures Specific to Sporozoites and Oocysts Heather M. Fritz, Kerry R. Buchholz, Xiucui Chen, Blythe Durbin-Johnson, David M. Rocke, Patricia A. Conrad, John C. Boothroyd, Silvia N. Moreno PLoS ONE.2012; 7(2): e29998. CrossRef
Toxoplasma gondii: Expression of GRA1 gene in endoplasmic reticulum promotes both growth and adherence and modulates intracellular calcium release in macrophages Jie Lin, Xu Lin, Guo-hua Yang, Yong Wang, Bi-wen Peng, Jian-yin Lin Experimental Parasitology.2010; 125(2): 165. CrossRef
Schizophrenia Susceptibility Genes Directly Implicated in the Life Cycles of Pathogens: Cytomegalovirus, Influenza, Herpes simplex, Rubella, and Toxoplasma gondii C.J. Carter Schizophrenia Bulletin.2009; 35(6): 1163. CrossRef
GRA Proteins of Toxoplasma gondii: Maintenance of Host-Parasite Interactions across the Parasitophorous Vacuolar Membrane Ho-Woo Nam The Korean Journal of Parasitology.2009; 47(Suppl): S29. CrossRef
Interaction between Parasitophorous Vacuolar Membrane-associated GRA3 and Calcium Modulating Ligand of Host Cell Endoplasmic Reticulum in the Parasitism of Toxoplasma gondii Ji Yeon Kim, Hye-Jin Ahn, Kyung Ju Ryu, Ho-Woo Nam The Korean Journal of Parasitology.2008; 46(4): 209. CrossRef
Nucleolar translocalization of GRA10 of Toxoplasma gondii transfectionally expressed in HeLa cells Hye-Jin Ahn, Sehra Kim, Ho-Woo Nam The Korean Journal of Parasitology.2007; 45(3): 165. CrossRef
First
Prev
