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:	Salton GD; a Post-Graduation Program in Cellular and Molecular Biology, Molecular Radiobiology Laboratory, Biotechnology Center , Universidade Federal do Rio Grande do Sul , Porto Alegre (RS) , Brazil , Cryobiology Unit and Umbilical Cord Blood Bank, Hemotherapy Service , Hospital de Clínicas de Porto Alegre , Porto Alegre (RS) , Brazil., Laurino CCFC; b Molecular Biology for Auto-immune and Infectious Diseases Laboratory, Experimental Research Center, Hospital de Clínicas de Porto Alegre , Universidade Federal do Rio Grande do Sul , Porto Alegre (RS) , Brazil . Embriology and Cellular Differentiation Laboratory, Experimental Research Center, Hospital de Clínicas de Porto Alegre; Faculdade de Veterinária , Universidade Federal do Rio Grande do Sul , Porto Alegre (RS) , Brazil . Faculdade Nossa Senhora de Fátima , Caxias do Sul (RS) , Brazil . Instituto Brasileiro de Saúde , Porto Alegre (RS) , Brazil., Mega NO; c Animal Biology Post-Graduation Program , Universidade Federal do Rio Grande do Sul , Porto Alegre (RS) , Brazil., Delgado-Cañedo A; d Biotechnology Research Center for Interdisciplinary Research , Universidade Federal do Pampa , São Gabriel (RS) , Brazil., Setterblad N; e Imaging, Cell Selection and Genomics Platform , Institut Universitaire d'Hématologie, Hôpital Saint-Louis , Paris , France., Carmagnat M; f Immunology and Histocompatibility Laboratory AP-HP , INSERM UMRS 940, Institut Universitaire d'Hématologie , Paris , France., Xavier RM; g Molecular Biology for Auto-immune and Infectious Diseases Laboratory, Experimental Research Center, Hospital de Clínicas de Porto Alegre , Universidade Federal do Rio Grande do Sul , Porto Alegre (RS) , Brazil., Cirne-Lima E; h Embriology and Cellular Differentiation Laboratory, Experimental Research Center, Hospital de Clínicas de Porto Alegre; Faculdade de Veterinária , Universidade Federal do Rio Grande do Sul , Porto Alegre (RS) , Brazil., Lenz G; i Cell Signaling Laboratory, Biophysics Department, Biotechnology Center and Post-Graduation Program in Cellular and Molecular Biology , Universidade Federal do Rio Grande do Sul , Porto Alegre (RS) , Brazil., Henriques JAP; j Molecular Radiobiology Laboratory, Biotechnology Center and Post-Graduation Program in Cellular and Molecular Biology , Universidade Federal do Rio Grande do Sul , Porto Alegre (RS) ; Biotechnology Institute , Universidade de Caxias do Sul , Caxias do Sul (RS) , Brazil., Laurino JP; k Biotechnology Institute , Universidade de Caxias do Sul, Caxias do Sul (RS) and Instituto Brasileiro de Saúde , Porto Alegre (RS) , Brazil.
Jazyk:	angličtina
Zdroj:	Cancer biology & therapy [Cancer Biol Ther] 2017 Aug 03; Vol. 18 (8), pp. 560-570. Date of Electronic Publication: 2017 Jul 10.
DOI:	10.1080/15384047.2017.1345383
Abstrakt:	Background: 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. Methods: 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. Results: 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. Discussion: 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:	MEDLINE
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