title: Identification of Cyclobutane Pyrimidine Dimer-Responsive Genes Using UVB-Irradiated Human Keratinocytes Transfected with In Vitro-Synthesized Photolyase mRNA.
abstract: Major biological effects of UVB are attributed to cyclobutane pyrimidine dimers (CPDs), the most common photolesions formed on DNA. To investigate the contribution of CPDs to UVB-induced changes of gene expression, a model system was established by transfecting keratinocytes with pseudouridine-modified mRNA (Ψ-mRNA) encoding CPD-photolyase. Microarray analyses of this model system demonstrated that more than 50% of the gene expression altered by UVB was mediated by CPD photolesions. Functional classification of the gene targets revealed strong effects of CPDs on the regulation of the cell cycle and transcriptional machineries. To confirm the microarray data, cell cycle-regulatory genes, CCNE1 and CDKN2B that were induced exclusively by CPDs were selected for further investigation. Following UVB irradiation, expression of these genes increased significantly at both mRNA and protein levels, but not in cells transfected with CPD-photolyase Ψ-mRNA and exposed to photoreactivating light. Treatment of cells with inhibitors of c-Jun N-terminal kinase (JNK) blocked the UVB-dependent upregulation of both genes suggesting a role for JNK in relaying the signal of UVB-induced CPDs into transcriptional responses. Thus, photolyase mRNA-based experimental platform demonstrates CPD-dependent and -independent events of UVB-induced cellular responses, and, as such, has the potential to identify novel molecular targets for treatment of UVB-mediated skin diseases.
authors.affiliation: Department of Dermatology, Faculty of Medicine, University of Debrecen, Debrecen, Hungary.
authors.forename: Gábor
authors.initials: G
authors.lastname: Boros
authors.affiliation: Department of Dermatology, Faculty of Medicine, University of Debrecen, Debrecen, Hungary.
authors.forename: Edit
authors.initials: E
authors.lastname: Miko
authors.affiliation: Department of Neurosurgery, University of Pennsylvania, Philadelphia, Pennsylvania, United States of America.
authors.forename: Hiromi
authors.initials: H
authors.lastname: Muramatsu
authors.affiliation: Department of Medicine, University of Pennsylvania, Philadelphia, Pennsylvania, United States of America.
authors.forename: Drew
authors.initials: D
authors.lastname: Weissman
authors.affiliation: Department of Dermatology, Faculty of Medicine, University of Debrecen, Debrecen, Hungary.
authors.forename: Eszter
authors.initials: E
authors.lastname: Emri
authors.affiliation: Department of Genetics, Erasmus University Medical Center, Rotterdam, The Netherlands.
authors.forename: Gijsbertus T J
authors.initials: GT
authors.lastname: van der Horst
authors.affiliation: Department of Dermatology, Faculty of Medicine, University of Debrecen, Debrecen, Hungary; Department of Dermatological Allergology, Faculty of Medicine, University of Debrecen, Debrecen, Hungary.
authors.forename: Andrea
authors.initials: A
authors.lastname: Szegedi
authors.affiliation: Department of Dermatology, Faculty of Medicine, University of Debrecen, Debrecen, Hungary.
authors.forename: Irén
authors.initials: I
authors.lastname: Horkay
authors.affiliation: Department of Dermatology, Faculty of Medicine, University of Debrecen, Debrecen, Hungary.
authors.forename: Gabriella
authors.initials: G
authors.lastname: Emri
authors.affiliation: Department of Neurosurgery, University of Pennsylvania, Philadelphia, Pennsylvania, United States of America.
authors.forename: Katalin
authors.initials: K
authors.lastname: Karikó
authors.affiliation: Department of Dermatology, Faculty of Medicine, University of Debrecen, Debrecen, Hungary.
authors.forename: Éva
authors.initials: É
authors.lastname: Remenyik
mesh_terms: D000818:Animals; D002460:Cell Line; D019927:Cyclin E / Q000235:genetics / Q000378:metabolism; D050762:Cyclin-Dependent Kinase Inhibitor p15 / Q000235:genetics / Q000378:metabolism; D004260:DNA Repair / Q000528:radiation effects; D004255:Deoxyribodipyrimidine Photo-Lyase / Q000235:genetics* / Q000378:metabolism; D005786:Gene Expression Regulation / Q000528:radiation effects*; D006801:Humans; D048031:JNK Mitogen-Activated Protein Kinases / Q000378:metabolism; D015603:Keratinocytes / Q000201:enzymology / Q000378:metabolism* / Q000528:radiation effects; D020935:MAP Kinase Signaling System / Q000528:radiation effects; D020411:Oligonucleotide Array Sequence Analysis; D015513:Oncogene Proteins / Q000235:genetics / Q000378:metabolism; D048328:Potoroidae; D011740:Pyrimidine Dimers / Q000378:metabolism*; D012333:RNA, Messenger / Q000235:genetics / Q000378:metabolism; D060888:Real-Time Polymerase Chain Reaction; D015203:Reproducibility of Results; D013312:Stress, Physiological / Q000528:radiation effects; D014158:Transcription, Genetic / Q000528:radiation effects; D014162:Transfection; D014466:Ultraviolet Rays
chemical_list: C475820:CCNE1 protein, human; C493966:CDKN2B protein, human; D019927:Cyclin E; D050762:Cyclin-Dependent Kinase Inhibitor p15; D015513:Oncogene Proteins; D011740:Pyrimidine Dimers; D012333:RNA, Messenger; D048031:JNK Mitogen-Activated Protein Kinases; D004255:Deoxyribodipyrimidine Photo-Lyase