Differential roles of p63 isoforms in epidermal development: selective genetic complementation in p63 null mice (original) (raw)

Cell Death & Differentiation volume 13, pages 1037–1047 (2006)Cite this article

Abstract

Epidermal development requires the transcription factor p63, as p63−/− mice are born dead, without skin. The gene expresses two proteins, one with an amino-terminal transactivation domain (TAp63) and one without (ΔNp63), although their relative contribution to epidermal development is unknown. To address this issue, we reintroduced TAp63_α_ and/or ΔNp63_α_ under the K5 promoter into p63−/− mice by in vivo genetic complementation. Whereas p63−/− and p63−/−;TA mice showed extremely rare patches of poorly differentiated keratinocytes, p63−/−;ΔN mice showed significant epidermal basal layer formation. Double TAp63_α_/ΔNp63_α_ complementation showed greater patches of differentiated skin; at the ultrastructural level, there was clear reformation of a distinct basal membrane and hemidesmosomes. At the molecular level, ΔNp63 regulated expression of genes characteristic of the basal layer (K14), interacting (by Chip, luc assay) with the third p53 consensus site. Conversely, TAp63 transcribed the upper layer's genes (Ets-1, K1, transglutaminases, involucrin). Therefore, the two p63 isoforms appear to play distinct cooperative roles in epidermal formation.

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Abbreviations

TA:

transactivation domain

ΔN:

amino-terminal truncated protein

H&E:

haematoxylin and eosin

tg:

transgenic mice

p63−/−;ΔN:

mice knockout for p63, re-expressing ΔNp63_α_

p63−/−;TA:

mice knockout for p63, re-expressing TAp63_α_

p63−/−;ΔN;TA:

mice knockout for p63, re-expressing both ΔNp63_α_ and TAp73_α_

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Acknowledgements

We thank Dr. F McKeon for providing us with the p63−/− mice, Dr. C Tiveron for crucial help and advice in the generation of the transgenic mice, and Mr. A Colangeli and J McWilliam for ultramicrotomy. This work was supported by grants from Telethon to EC; AIRC to VDL; and EU, AIRC, Telethon, FIRB, MIUR, MinSan, TelethonGGP04110 and MRC to GM.

Author information

Authors and Affiliations

  1. Biochemistry Laboratory, IDI-IRCCS, c/o University of Rome ‘Tor Vergata’, Rome, 00133, Italy
    E Candi, A Rufini, A Terrinoni, M Ranalli, A Paradisi, L G Spagnoli, M V Catani, S Ramadan & G Melino
  2. Medical Research Council, Toxicology Unit, Leicester University, Leicester, LE1 9HN, UK
    D Dinsdale, V De Laurenzi, R A Knight & G Melino

Authors

  1. E Candi
  2. A Rufini
  3. A Terrinoni
  4. D Dinsdale
  5. M Ranalli
  6. A Paradisi
  7. V De Laurenzi
  8. L G Spagnoli
  9. M V Catani
  10. S Ramadan
  11. R A Knight
  12. G Melino

Corresponding author

Correspondence toG Melino.

Additional information

Edited by P Vandenabeele

Note added in proof

After this paper was accepted, Laurikkala et al. (Laurikkala J, Mikkola ML, James M, Tummers M, Mills AA and Thesleff I (2006) P63 regulates multiple signaling pathways required for ectodermal organogenesis and differentiation. Development 133: in press. Published online on March 8, 2006. doi: 10.1242/dev.0235) demonstrated that ΔNp63 is the main isoform expressed at all embryonic stages during epidermal, tooth and hair development, accounting for 100% of all p63 expressed up to E9 and 99% at E13. TAp63 expression starts at E13, and only accounts for 1% of total p63 protein expressed at this time. Thus, expression of the ΔNp63 isoform is predominant over that of TAp63, at a time before any epidermal stratification occurs. These results obtained by Laurikkala et al. are fully in agreement with our studies, using a completely different experimental approach, and confirm that the ΔNp63 isoform is crucial for the formation of the epidermis, whereas TAp63 contributes to the control of epithelial differentiation by acting synergistically and/or subsequently to ΔNp63.

Moreover, Laurikkala et al. also demonstrated that Bmp 7, Fgfr2b, Jag1 and Notch1 transcripts are coexpressed with ΔNp63 and are absent in p63−/−mice. Additional support for this complex pathway of regulations also comes from a further independent paper in press (Nguyen BC, Lefort K, Mandinova A, Antonini D, Devgan V, Della Gatta G, Koster MI, Zhang Z, Wang J, Tommasi di Vignano A, Kitajewski J, Chiorino G, Roop DR, Missero C and Dotto GP (2006). Cross- regulation between Notch and p63 in keratinocyte commitment to differentiation. Genes Dev. in press. doi:10.1101/gad1406006).

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Candi, E., Rufini, A., Terrinoni, A. et al. Differential roles of p63 isoforms in epidermal development: selective genetic complementation in p63 null mice.Cell Death Differ 13, 1037–1047 (2006). https://doi.org/10.1038/sj.cdd.4401926

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