CPD-photolyase adenovirus-mediated gene transfer in normal and DNA-repair-deficient human cells

Journal of Cell Science
Vanessa ChigançasCarlos F M Menck

Abstract

Cyclobutane pyrimidine dimers (CPDs) are the most frequent and deleterious lesions generated in the mammalian genome after UV-C irradiation. The persistence of these lesions in DNA can be toxic and mutagenic, and also represents a specific signal to apoptosis. To investigate the CPDs repair in situ and consequent UV-induced apoptosis in human cells, we generated a recombinant adenovirus vector containing the gene encoding a CPD-photolyase-EGFP fusion protein (Adphr-EGFP). Adphr-EGFP-infected cells are proficient in photorepair, which prevents apoptotic cell death in comparison with samples kept in the dark, indicating that the fusion protein is functional in CPD recognition and removal. By using local UV irradiation, foci of the photolyase fusion protein were observed in UV-damaged areas of the nuclei in colocalization with NER enzymes. Phr-EGFP migration to CPD sites and redistribution after photorepair was followed, and shown to present similar kinetics in normal or DNA-repair-deficient cells. To our knowledge, this is the first report of an investigation of CPDs repair in situ employing a CPD-photolyase-EGFP enzyme. The Adphr-EGFP vector can be an informative tool to investigate the repair and cellular consequences of UV-ind...Continue Reading

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Citations

May 17, 2006·Apoptosis : an International Journal on Programmed Cell Death·Luis Francisco Zirnberger BatistaCarlos Frederico Martins Menck
Dec 22, 2007·DNA Repair·Keronninn Moreno de Lima-BessaCarlos Frederico Martins Menck
Jun 27, 2006·DNA Repair·Keronninn Moreno Lima-BessaCarlos Frederico Martins Menck
Dec 20, 2012·Pharmaceutical Biology·Marioly VernhesAngel Sánchez-Lamar
Mar 21, 2015·Journal of Invertebrate Pathology·Qi TangGuohui Li
Nov 29, 2007·Human & Experimental Toxicology·Melissa G ArmeliniCarlos F M Menck
Apr 18, 2006·Biochemistry. Biokhimii︠a︡·E G NovoselovaE E Fesenko
Jul 5, 2013·Photochemical & Photobiological Sciences : Official Journal of the European Photochemistry Association and the European Society for Photobiology·Barbara CortatCarlos Frederico Martins Menck
May 24, 2020·Proceedings of the National Academy of Sciences of the United States of America·Kai-Feng HungMasaoki Kawasumi

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Apoptosis

Apoptosis is a specific process that leads to programmed cell death through the activation of an evolutionary conserved intracellular pathway leading to pathognomic cellular changes distinct from cellular necrosis