Defective transcription of ATF3 responsive genes, a marker for Cockayne Syndrome

• Published in: Scientific reports 2020-01, Vol.10 (1), p.1105-1105, Article 1105
• Main Authors: Epanchintsev, Alexey, Rauschendorf, Marc-Alexander, Costanzo, Federico, Calmels, Nadege, Obringer, Cathy, Sarasin, Alain, Coin, Frederic, Laugel, Vincent, Egly, Jean-Marc
• Format: Article
• Language: English
• Subjects: 38/39; 38/91; 42/41; 631/208/199; 631/337/1427/1429; 631/337/572; Activating transcription factor 3; Activating Transcription Factor 3 - genetics; Activating Transcription Factor 3 - metabolism; Biochemistry, Molecular Biology; Cell Cycle Proteins; Cell Line; Cell lines; Chromatin; Cockayne syndrome; Cockayne Syndrome - diagnosis; Cockayne Syndrome - genetics; DNA Damage; DNA Helicases - genetics; DNA Repair Enzymes - genetics; Gene expression; Gene Expression Profiling; Genes, Immediate-Early - genetics; Genetic disorders; Genetic Markers; Genetics; Genotoxicity; Human genetics; Humanities and Social Sciences; Humans; Immunofluorescence; Life Sciences; Molecular biology; multidisciplinary; Mutation; Nerve Tissue Proteins; Neuregulin-1; Photosensitivity; Poly-ADP-Ribose Binding Proteins - genetics; Polymerase chain reaction; Regulatory sequences; RNA Polymerase II - metabolism; Science; Science (multidisciplinary); Transcription; Transcription Factors - genetics; Transcription, Genetic - genetics; Ultraviolet Rays
• ISSN: 2045-2322; 2045-2322
• DOI: 10.1038/s41598-020-57999-4

Cockayne syndrome (CS) is a rare genetic disorder caused by mutations (dysfunction) in CSA and CSB . CS patients exhibit mild photosensitivity and severe neurological problems. Currently, CS diagnosis is based on the inefficiency of CS cells to recover RNA synthesis upon genotoxic (UV) stress. Indeed, upon genotoxic stress, ATF3 , an immediate early gene is activated to repress up to 5000 genes encompassing its responsive element for a short period of time. On the contrary in CS cells, CSA and CSB dysfunction impairs the degradation of the chromatin-bound ATF3, leading to a permanent transcriptional arrest as observed by immunofluorescence and ChIP followed by RT-PCR. We analysed ChIP-seq of Pol II and ATF3 promoter occupation analysis and RNA sequencing-based gene expression profiling in CS cells, as well as performed immunofluorescence study of ATF3 protein stability and quantitative RT-PCR screening in 64 patient cell lines. We show that the analysis of few amount (as for example CDK5RAP2, NIPBL and NRG1 ) of ATF3 dependent genes, could serve as prominent molecular markers to discriminate between CS and non-CS patient’s cells. Such assay can significantly simplify the timing and the complexity of the CS diagnostic procedure in comparison to the currently available methods.