Eimeria tenella: 14-3-3 protein interacts with telomerase

Area of Science:

• Molecular Biology
• Parasitology
• Biochemistry

Background:

• Telomerase maintains chromosome ends, with telomerase reverse transcriptase (TERT) as its catalytic subunit.
• TERT activity is regulated by associated proteins involved in assembly, modification, localization, and function.

Purpose of the Study:

• To investigate protein interactions with the telomerase RNA-binding domain of Eimeria tenella TERT (EtTRBD).
• To identify proteins that associate with EtTRBD and elucidate their role in telomerase regulation.

Main Methods:

• Cloning of EtTRBD into a yeast two-hybrid vector (pGBKT7) to serve as bait.
• Yeast two-hybrid screening to identify interacting proteins.
• Prokaryotic and eukaryotic expression of EtTRBD and the identified interacting protein (14-3-3).
• Pull-down assays and co-immunoprecipitation to confirm protein-protein interactions.

Main Results:

• The EtTRBD bait plasmid did not self-activate the Gal4 reporter gene.
• The 14-3-3 protein was identified as an EtTRBD-associated protein via yeast two-hybrid screening.
• Pull-down assays confirmed a direct binding interaction between purified EtTRBD and 14-3-3.
• Co-immunoprecipitation validated the interaction between EtTRBD and 14-3-3 in 293T cells.

Conclusions:

• The 14-3-3 protein directly interacts with the telomerase RNA-binding domain of Eimeria tenella TERT.
• This protein-protein interaction offers a foundation for further research into telomerase and telomere regulation in E. tenella.