title: Eimeria tenella: 14-3-3 protein interacts with telomerase.
abstract: Telomerase, consisting of telomerase RNA and telomerase reverse transcriptase (TERT), is responsible for the maintenance of the end of linear chromosomes. TERT, as the catalytic subunit of telomerase, plays a critical role in telomerase activity. Researches indicate TERT-associated proteins participate in the regulation of telomerase assembly, posttranslational modification, localization, and enzymatic function. Here, the telomerase RNA-binding domain of Eimeria tenella TERT (EtTRBD) was cloned into pGBKT7 and performed as the bait. α-Galactosidase assay showed that the bait plasmid did not activate Gal4 reporter gene. Further, we isolated an EtTRBD-associated protein, 14-3-3, by yeast two-hybrid screening using the constructed bait plasmid. To confirm the interaction, EtTRBD and 14-3-3 were expressed by prokaryotic and eukaryotic expression systems. Pull-down assays by purified proteins demonstrated a direct bind between EtTRBD and 14-3-3. Co-immunoprecipitation techniques successfully validated that 14-3-3 interacted with EtTRBD in 293T cells. The protein-protein interaction provides a starting point for more in-depth studies on telomerase and telomere regulation in E. tenella.
authors.affiliation: Key Laboratory of Zoonosis Research, Ministry of Education, College of Veterinary Medicine, Jilin University, 5333 Xi'an Road, Changchun, 130062, China.
authors.forename: Na
authors.initials: N
authors.lastname: Zhao
authors.forename: Pengtao
authors.initials: P
authors.lastname: Gong
authors.forename: Baiqi
authors.initials: B
authors.lastname: Cheng
authors.forename: Jianhua
authors.initials: J
authors.lastname: Li
authors.forename: Zhengtao
authors.initials: Z
authors.lastname: Yang
authors.forename: He
authors.initials: H
authors.lastname: Li
authors.forename: Ju
authors.initials: J
authors.lastname: Yang
authors.forename: Guocai
authors.initials: G
authors.lastname: Zhang
authors.forename: Xichen
authors.initials: X
authors.lastname: Zhang
mesh_terms: D048948:14-3-3 Proteins / Q000235:genetics / Q000378:metabolism*; D016786:Eimeria tenella / Q000235:genetics / Q000378:metabolism*; D047468:Immunoprecipitation; D010957:Plasmids; D011485:Protein Binding; D054730:Protein Interaction Domains and Motifs; D015800:Protozoan Proteins / Q000235:genetics / Q000378:metabolism*; D012313:RNA; D011994:Recombinant Proteins / Q000378:metabolism; D019098:Telomerase / Q000235:genetics / Q000378:metabolism*; D020798:Two-Hybrid System Techniques
chemical_list: D048948:14-3-3 Proteins; D015800:Protozoan Proteins; D011994:Recombinant Proteins; C422952:telomerase RNA; D012313:RNA; D019098:Telomerase