Identification of host proteins interacting with Toxoplasma gondii GRA15 (TgGRA15) by yeast two-hybrid system
Background Toxoplasma gondii, an obligate intracellular protozoan parasite, possesses the remarkable ability to co-opt host cell machinery in order to maintain its intracellular survival. This parasite can modulate signaling pathways of its host through the secretion of polymorphic effector prote...
| Main Authors: | , , , , |
|---|---|
| Format: | Article |
| Published: |
BioMed Central
2017
|
| Subjects: | |
| Online Access: | https://eprints.nottingham.ac.uk/40339/ |
| _version_ | 1848796032805109760 |
|---|---|
| author | Liu, Qing Li, Fa-Cai Elsheikha, Hany M. Sun, Miao-Miao Zhu, Xing-Quan |
| author_facet | Liu, Qing Li, Fa-Cai Elsheikha, Hany M. Sun, Miao-Miao Zhu, Xing-Quan |
| author_sort | Liu, Qing |
| building | Nottingham Research Data Repository |
| collection | Online Access |
| description | Background
Toxoplasma gondii, an obligate intracellular protozoan parasite, possesses the remarkable ability to co-opt host cell machinery in order to maintain its intracellular survival. This parasite can modulate signaling pathways of its host through the secretion of polymorphic effector proteins localized in the rhoptry and dense granule organelles. One of such effectors is T. gondii type II-specific dense granule protein 15, TgGRA15, which activates NF-κB pathway. The aim of the present study was to identify the host interaction partner proteins of TgGRA15.
Methods
We screened a yeast two-hybrid mouse cDNA library using TgGRA15 as the bait. TgGRA15 (PRU strain, Type II) was cloned into the pGBKT7 vector and expressed in the Y2HGold yeast strain. Then, the bait protein expression was validated by western blotting analysis, followed by auto-activation and toxicity tests in comparison with control (Y2HGold yeast strain transformed with empty pGBKT7 vector).
Results
This screening led to the identification of mouse Luzp1 and AW209491 as host binding proteins that interact with TgGRA15. Luzp1 contains three nuclear localizing signals and is involved in regulating a subset of host non-coding RNA genes.
Conclusions
These findings reveal, for the first time, new host cell proteins interacting with TgGRA15. The identification of these cellular targets and the understanding of their contribution to the host-pathogen interaction may serve as the foundation for novel therapeutic and prevention strategies against T. gondii infection. |
| first_indexed | 2025-11-14T19:41:32Z |
| format | Article |
| id | nottingham-40339 |
| institution | University of Nottingham Malaysia Campus |
| institution_category | Local University |
| last_indexed | 2025-11-14T19:41:32Z |
| publishDate | 2017 |
| publisher | BioMed Central |
| recordtype | eprints |
| repository_type | Digital Repository |
| spelling | nottingham-403392020-05-04T18:31:56Z https://eprints.nottingham.ac.uk/40339/ Identification of host proteins interacting with Toxoplasma gondii GRA15 (TgGRA15) by yeast two-hybrid system Liu, Qing Li, Fa-Cai Elsheikha, Hany M. Sun, Miao-Miao Zhu, Xing-Quan Background Toxoplasma gondii, an obligate intracellular protozoan parasite, possesses the remarkable ability to co-opt host cell machinery in order to maintain its intracellular survival. This parasite can modulate signaling pathways of its host through the secretion of polymorphic effector proteins localized in the rhoptry and dense granule organelles. One of such effectors is T. gondii type II-specific dense granule protein 15, TgGRA15, which activates NF-κB pathway. The aim of the present study was to identify the host interaction partner proteins of TgGRA15. Methods We screened a yeast two-hybrid mouse cDNA library using TgGRA15 as the bait. TgGRA15 (PRU strain, Type II) was cloned into the pGBKT7 vector and expressed in the Y2HGold yeast strain. Then, the bait protein expression was validated by western blotting analysis, followed by auto-activation and toxicity tests in comparison with control (Y2HGold yeast strain transformed with empty pGBKT7 vector). Results This screening led to the identification of mouse Luzp1 and AW209491 as host binding proteins that interact with TgGRA15. Luzp1 contains three nuclear localizing signals and is involved in regulating a subset of host non-coding RNA genes. Conclusions These findings reveal, for the first time, new host cell proteins interacting with TgGRA15. The identification of these cellular targets and the understanding of their contribution to the host-pathogen interaction may serve as the foundation for novel therapeutic and prevention strategies against T. gondii infection. BioMed Central 2017-01-03 Article PeerReviewed Liu, Qing, Li, Fa-Cai, Elsheikha, Hany M., Sun, Miao-Miao and Zhu, Xing-Quan (2017) Identification of host proteins interacting with Toxoplasma gondii GRA15 (TgGRA15) by yeast two-hybrid system. Parasites & Vectors, 10 (1). ISSN 1756-3305 Toxoplasma gondii TgGRA15 Host-pathogen interaction Yeast-two-hybrid Luzp1 AW209491 http://parasitesandvectors.biomedcentral.com/articles/10.1186/s13071-016-1943-1 doi:10.1186/s13071-016-1943-1 doi:10.1186/s13071-016-1943-1 |
| spellingShingle | Toxoplasma gondii TgGRA15 Host-pathogen interaction Yeast-two-hybrid Luzp1 AW209491 Liu, Qing Li, Fa-Cai Elsheikha, Hany M. Sun, Miao-Miao Zhu, Xing-Quan Identification of host proteins interacting with Toxoplasma gondii GRA15 (TgGRA15) by yeast two-hybrid system |
| title | Identification of host proteins interacting with Toxoplasma gondii GRA15 (TgGRA15) by yeast two-hybrid system |
| title_full | Identification of host proteins interacting with Toxoplasma gondii GRA15 (TgGRA15) by yeast two-hybrid system |
| title_fullStr | Identification of host proteins interacting with Toxoplasma gondii GRA15 (TgGRA15) by yeast two-hybrid system |
| title_full_unstemmed | Identification of host proteins interacting with Toxoplasma gondii GRA15 (TgGRA15) by yeast two-hybrid system |
| title_short | Identification of host proteins interacting with Toxoplasma gondii GRA15 (TgGRA15) by yeast two-hybrid system |
| title_sort | identification of host proteins interacting with toxoplasma gondii gra15 (tggra15) by yeast two-hybrid system |
| topic | Toxoplasma gondii TgGRA15 Host-pathogen interaction Yeast-two-hybrid Luzp1 AW209491 |
| url | https://eprints.nottingham.ac.uk/40339/ https://eprints.nottingham.ac.uk/40339/ https://eprints.nottingham.ac.uk/40339/ |