Proteome alteration induced by hTERT transfection of human fibroblast cells.

Proteome Science
Gabriel D MazzucchelliE De Pauw

Abstract

Telomerase confers cellular immortality by elongating telomeres, thereby circumventing the Hayflick limit. Extended-life-span cells have been generated by transfection with the human telomerase reverse transcriptase (hTERT) gene. hTERT transfected cell lines may be of outstanding interest to monitor the effect of drugs targeting the telomerase activity. The incidence of hTERT gene transfection at the proteome level is a prerequisite to that purpose. The effect of the transfection has been studied on the proteome of human fibroblast (WI38). Cytosolic and nuclear fractions of WI38 cells, empty vector transfected WI38 (WI38-HPV) and hTERT WI38 cells were submitted to a 2D-DIGE (Two-Dimensional Differential In-Gel Electrophoresis) analysis. Only spots that had a similar abundance in WI38 and WI38-HPV, but were differentially expressed in WI38 hTERT were selected for MS identification. This method directly points to the proteins linked with the hTERT expression. Number of false positive differentially expressed proteins has been excluded by using control WI38-HPV cells. The proteome alteration induced by hTERT WI38 transfection should be taken into account in subsequent use of the cell line for anti-telomerase drugs evaluation. 2D-D...Continue Reading

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Citations

Mar 5, 2009·BioEssays : News and Reviews in Molecular, Cellular and Developmental Biology·Bent Honoré
Jul 9, 2013·International Journal of Medical Microbiology : IJMM·Inken PadbergThomas F Meyer
Sep 20, 2016·Journal of the American Chemical Society·Hyun-Jin KangLaurence H Hurley
Apr 15, 2009·Journal of Proteome Research·Jing ShenYingnian Yu
May 1, 2021·International Journal of Molecular Sciences·Nadezhda A EvtushenkoNadya G Gurskaya

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Methods Mentioned

BETA
transfection
FACS
electrophoresis
interactions predictions
gene knockout
protein folding
flow cytometry

Software Mentioned

Mascot
Biotools
STRING

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