Selective DNA damage responses in murine Xpa-/-, Xpc-/- and Csb-/- keratinocyte cultures

Gerdine J Stout, Marijke van Oosten, Fatima Z Acherrat, Jan de Wit, Wilbert P Vermeij, Leon H F Mullenders, Frank R de Gruijl, Claude Backendorf

Research output: Contribution to journalArticlepeer-review

14 Citations (Scopus)


Cellular DNA damage responses (DDRs) are induced by unrepaired DNA lesions and constitute a protective back-up system that prevents the expansion of damaged cells. These cellular signaling pathways trigger either growth arrest or cell death and are believed to be major components of an early anti-cancer barrier. Cultures of C57BL/6J keratinocytes with various defects in NER sub-pathways allowed us to follow the kinetics of DDRs in an isogenic background and in the proper (physiologically relevant) target cells, supplementing earlier studies in heterogenic human fibroblasts. In a series of well-controlled parallel experiments we have shown that, depending on the NER deficiency, murine keratinocytes elicited highly selective DDRs. After a dose of UV-B that did not affect wild-type keratinocytes, Xpa(-/-) keratinocytes (complete NER deficiency) showed a rapid depletion of DNA replicating S-phase cells, a transient increase in quiescent S-phase cells (not replicating DNA), followed by massive apoptosis. Csb(-/-) keratinocytes (TC-NER deficient) responded by a more sustained increase in QS-phase cells and appeared more resistant to UV-B induced apoptosis than Xpa(-/-). In irradiated Xpc(-/-) keratinocytes (GG-NER deficient) the loss of replicating S-phase cells was associated with a gradual build-up of both QS-phase cells and cells arrested in late-S phase, in complete absence of apoptosis. Our analysis complements and extends previous in vivo investigations and highlights both similarities and differences with earlier fibroblast studies. In vitro cultures of murine keratinocytes provide a new tool to unravel the molecular mechanisms of UV-induced cellular stress responses in great detail and in a physiologically relevant background. This will be essential to fully appreciate the implications of DDRs in tumor suppression and cancer prevention.

Original languageEnglish
Pages (from-to)1337-44
Number of pages8
JournalDNA repair
Issue number11
Publication statusPublished - 21 Nov 2005
Externally publishedYes


  • Animals
  • Apoptosis/genetics
  • Cell Death/radiation effects
  • Cell Division/radiation effects
  • Cells, Cultured
  • DNA Damage/genetics
  • DNA Repair/genetics
  • DNA Repair Enzymes/genetics
  • DNA Replication/radiation effects
  • DNA-Binding Proteins/genetics
  • Genetic Predisposition to Disease
  • Keratinocytes/metabolism
  • Mice
  • Poly-ADP-Ribose Binding Proteins
  • Radiation Tolerance
  • S Phase/genetics
  • Signal Transduction/genetics
  • Skin Neoplasms/etiology
  • Ultraviolet Rays
  • Xeroderma Pigmentosum Group A Protein/genetics


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