title: Restoring DNA repair capacity of cells from three distinct diseases by XPD gene-recombinant adenovirus.
abstract: The nucleotide excision repair (NER) is one of the major human DNA repair pathways. Defects in one of the proteins that act in this system result in three distinct autosomal recessive syndromes: xeroderma pigmentosum (XP), Cockayne syndrome (CS) and trichothiodystrophy (TTD). TFIIH is a nine-protein complex essential for NER activity, initiation of RNA polymerase II transcription and with a possible role in cell cycle regulation. XPD is part of the TFIIH complex and has a helicase function, unwinding the DNA in the 5' --> 3' direction. Mutations in the XPD gene are found in XP, TTD and XP/CS patients, the latter exhibiting both XP and CS symptoms. Correction of DNA repair defects of these cells by transducing the complementing wild-type gene is one potential strategy for helping these patients. Over the last years, adenovirus vectors have been largely used in gene delivering because of their efficient transduction, high titer, and stability. In this work, we present the construction of a recombinant adenovirus carrying the XPD gene, which is coexpressed with the EGFP reporter gene by an IRES sequence, making it easier to follow cell infection. Infection by this recombinant adenovirus grants full correction of SV40-transformed and primary skin fibroblasts obtained from XP-D, TTD and XP/CS patients.
authors.affiliation: Department of Microbiology, Institute of Biomedical Sciences, University of São Paulo, São Paulo 05508-900 SP, Brazil.
authors.forename: Melissa Gava
authors.initials: MG
authors.lastname: Armelini
authors.forename: Alysson Renato
authors.initials: AR
authors.lastname: Muotri
authors.forename: Maria Carolina Nasser
authors.initials: MC
authors.lastname: Marchetto
authors.forename: Keronninn Moreno
authors.initials: KM
authors.lastname: de Lima-Bessa
authors.forename: Alain
authors.initials: A
authors.lastname: Sarasin
authors.forename: Carlos Frederico Martins
authors.initials: CF
authors.lastname: Menck
mesh_terms: D000256:Adenoviridae / Q000235:genetics*; D015153:Blotting, Western; D002460:Cell Line; D002461:Cell Line, Transformed; D045744:Cell Line, Tumor; D002469:Cell Separation; D002470:Cell Survival; D002478:Cells, Cultured; D003057:Cockayne Syndrome / Q000235:genetics / Q000628:therapy; D004247:DNA / Q000378:metabolism; D004265:DNA Helicases / Q000235:genetics*; D004260:DNA Repair; D004268:DNA-Binding Proteins / Q000235:genetics*; D004307:Dose-Response Relationship, Radiation; D004476:Ectodermal Dysplasia / Q000235:genetics / Q000628:therapy; D005347:Fibroblasts / Q000378:metabolism; D005434:Flow Cytometry; D005816:Genetic Complementation Test; D006801:Humans; D007700:Kinetics; D008297:Male; D008856:Microscopy, Fluorescence; D008957:Models, Genetic; D009154:Mutation; D010641:Phenotype; D012319:RNA Polymerase II / Q000378:metabolism; D011995:Recombination, Genetic; D012680:Sensitivity and Specificity; D012867:Skin / Q000378:metabolism; D051758:Transcription Factor TFIIH; D014157:Transcription Factors / Q000235:genetics*; D019473:Transcription Factors, TFII / Q000235:genetics; D014466:Ultraviolet Rays; D014983:Xeroderma Pigmentosum / Q000235:genetics / Q000628:therapy; D051759:Xeroderma Pigmentosum Group D Protein
chemical_list: D004268:DNA-Binding Proteins; D014157:Transcription Factors; D019473:Transcription Factors, TFII; D051758:Transcription Factor TFIIH; D004247:DNA; D012319:RNA Polymerase II; D004265:DNA Helicases; D051759:Xeroderma Pigmentosum Group D Protein; C497292:ERCC2 protein, human