Deletion of eIF2β lysine stretches creates a dominant negative that affects the translation and proliferation in human cell line: A tool for arresting the cell growth
| Autor: | Claudia Cilene Fernandes Correia Laurino, Elizabeth Obino Cirne-Lima, Gabrielle Dias Salton, Jomar Pereira Laurino, Andrés Delgado-Cañedo, Ricardo Machado Xavier, João Antonio Pêgas Henriques, Maryvonnick Carmagnat, Niclas Setterblad, Nicolás Oliveira Mega, Guido Lenz |
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| Rok vydání: | 2017 |
| Předmět: |
0301 basic medicine
Cytoplasm Cancer Research Cell Survival Apoptosis Biology 03 medical and health sciences chemistry.chemical_compound 0302 clinical medicine Neoplasms Humans Polylysine TetR Molecular Targeted Therapy RNA Messenger Cell Proliferation Sequence Deletion Cell Nucleus Pharmacology Zinc finger eIF2 Binding Sites Cell growth HEK 293 cells Cell sorting Molecular biology Cell biology G2 Phase Cell Cycle Checkpoints Eukaryotic Initiation Factor-2B Protein Transport HEK293 Cells 030104 developmental biology Oncology chemistry Protein Biosynthesis 030220 oncology & carcinogenesis Mutagenesis Site-Directed Molecular Medicine Protein Binding Research Paper |
| Zdroj: | Cancer Biology & Therapy. 18:560-570 |
| ISSN: | 1555-8576 1538-4047 |
| DOI: | 10.1080/15384047.2017.1345383 |
| Popis: | Eukaryote initiation factor 2 subunit β (eIF2β) plays a crucial role in regulation protein synthesis, which mediates the interaction of eIF2 with mRNA. eIF2β contains evolutionarily conserved polylysine stretches in amino-terminal region and a zinc finger motif in the carboxy-terminus.The gene eIF2β was cloned under tetracycline transcription control and the polylysine stretches were deleted by site-directed mutagenesis (eIF2βΔ3K). The plasmid was transfected into HEK 293 TetR cells. These cells were analyzed for their proliferative and translation capacities as well as cell death rate. Experiments were performed using gene reporter assays, western blotting, flow cytometry, cell sorting, cell proliferation assays and confocal immunofluorescence.eIF2βΔ3K affected negatively the protein synthesis, cell proliferation and cell survival causing G2 cell cycle arrest and increased cell death, acting in a negative dominant manner against the native protein. Polylysine stretches are also essential for eIF2β translocated from the cytoplasm to the nucleus, accumulating in the nucleolus and eIF2βΔ3K did not make this translocation.eIF2β is involved in the protein synthesis process and should act in nuclear processes as well. eIF2βΔ3K reduces cell proliferation and causes cell death. Since translation control is essential for normal cell function and survival, the development of drugs or molecules that inhibit translation has become of great interest in the scenario of proliferative disorders. In conclusion, our results suggest the dominant negative eIF2βΔ3K as a therapeutic strategy for the treatment of proliferative disorders and that eIF2β polylysine stretch domains are promising targets for this. |
| Databáze: | OpenAIRE |
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