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

Abstract

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.

Citation

Liu, Q., Li, F.-C., Elsheikha, H. M., Sun, M.-M., & Zhu, X.-Q. (2017). Identification of host proteins interacting with Toxoplasma gondii GRA15 (TgGRA15) by yeast two-hybrid system. Parasites and Vectors, 10(1), https://doi.org/10.1186/s13071-016-1943-1

• Journal Article Type: Article
• Acceptance Date: Dec 15, 2016
• Online Publication Date: Jan 3, 2017
• Publication Date: Jan 3, 2017
• Deposit Date: Feb 6, 2017
• Publicly Available Date: Feb 6, 2017
• Journal: Parasites & Vectors
• Electronic ISSN: 1756-3305
• Publisher: Springer Verlag
• Peer Reviewed: Peer Reviewed
• Volume: 10
• Issue: 1
• DOI: https://doi.org/10.1186/s13071-016-1943-1
• Keywords: Toxoplasma gondii, TgGRA15, Host-pathogen interaction, Yeast-two-hybrid, Luzp1, AW209491
• Public URL: https://nottingham-repository.worktribe.com/output/842203
• Publisher URL: http://parasitesandvectors.biomedcentral.com/articles/10.1186/s13071-016-1943-1
• Contract Date: Feb 6, 2017