Identification of cyclobutane pyrimidine dimer-responsive genes using UVB-irradiated human keratinocytes transfected with in vitro-synthesized photolyase mRNA

Gábor Boros, Edit Miko, Hiromi Muramatsu, Drew Weissman, Eszter Emri, Gijsbertus T J Van Der Horst, A. Szegedi, Irén Horkay, Gabriella Emri, Katalin Karikó, E. Remenyik

Research output: Article

4 Citations (Scopus)

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.

Original languageEnglish
Article numbere0131141
JournalPLoS One
Volume10
Issue number6
DOIs
Publication statusPublished - jún. 29 2015

Fingerprint

Deoxyribodipyrimidine Photo-Lyase
Pyrimidine Dimers
pyrimidines
keratinocytes
Keratinocytes
ultraviolet radiation
Genes
Messenger RNA
genes
Cells
Microarrays
Gene Expression
Gene expression
gene expression
cell cycle
Pseudouridine
In Vitro Techniques
cdc Genes
JNK Mitogen-Activated Protein Kinases
skin diseases

ASJC Scopus subject areas

  • Agricultural and Biological Sciences(all)
  • Biochemistry, Genetics and Molecular Biology(all)
  • Medicine(all)

Cite this

Identification of cyclobutane pyrimidine dimer-responsive genes using UVB-irradiated human keratinocytes transfected with in vitro-synthesized photolyase mRNA. / Boros, Gábor; Miko, Edit; Muramatsu, Hiromi; Weissman, Drew; Emri, Eszter; Van Der Horst, Gijsbertus T J; Szegedi, A.; Horkay, Irén; Emri, Gabriella; Karikó, Katalin; Remenyik, E.

In: PLoS One, Vol. 10, No. 6, e0131141, 29.06.2015.

Research output: Article

Boros, Gábor ; Miko, Edit ; Muramatsu, Hiromi ; Weissman, Drew ; Emri, Eszter ; Van Der Horst, Gijsbertus T J ; Szegedi, A. ; Horkay, Irén ; Emri, Gabriella ; Karikó, Katalin ; Remenyik, E. / Identification of cyclobutane pyrimidine dimer-responsive genes using UVB-irradiated human keratinocytes transfected with in vitro-synthesized photolyase mRNA. In: PLoS One. 2015 ; Vol. 10, No. 6.
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