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PMID: 17036050 Published · ppublish English Journal Article Research Support, Non-U.S. Gov't

New p63 targets in keratinocytes identified by a genome-wide approach.

The EMBO journal ·Vol. 25 ·No. 21 ·2006-11-01 ·Pages 5105-16

Viganò MA, Lamartine J, Testoni B, Merico D, Alotto D, Castagnoli C, Robert A, Candi E, Melino G, Gidrol X, Mantovani R

Abstract

p63 is a developmentally regulated transcription factor related to p53. It is involved in the development of ectodermal tissues, including limb, skin and in general, multilayered epithelia. The DeltaNp63alpha isoform is thought to play a 'master' role in the asymmetric division of epithelial cells. It is also involved in the pathogenesis of several human diseases, phenotypically characterized by ectodermal dysplasia. Our understanding of transcriptional networks controlled by p63 is limited, owing to the low number of bona fide targets. To screen for new targets, we employed chromatin immunoprecipitation from keratinocytes (KCs) coupled to the microarray technology, using both CpG islands and promoter arrays. The former revealed 96 loci, the latter yielded 85 additional genes. We tested 40 of these targets in several functional assays, including: (i) in vivo binding by p63 in primary KCs; (ii) expression analysis in differentiating HaCaT cells and in cells overexpressing DeltaNp63alpha; (iii) promoter transactivation and (iv) immunostaining in normal tissues, confirming their regulation by p63. We discovered several new specific targets whose functional categorization links p63 to cell growth and differentiation.

MeSH Terms
Animals COS Cells Cell Differentiation/physiology Chlorocebus aethiops DNA-Binding Proteins/genetics,metabolism Humans Keratinocytes/cytology,metabolism Organogenesis/physiology Protein Array Analysis/methods Protein Binding/physiology Trans-Activators/genetics,metabolism Transcription Factors Transcriptional Activation/physiology Tumor Suppressor Protein p53/metabolism Tumor Suppressor Proteins/genetics,metabolism
Chemicals
DNA-Binding Proteins TP63 protein, human Trans-Activators Transcription Factors Tumor Suppressor Protein p53 Tumor Suppressor Proteins
Authors & Affiliations
11 authors, click to expand affiliations / ORCID
Viganò M Alessandra
Dipartimento di Scienze Biomolecolari e Biotecnologie, Universita' di Milano, Milano, Italy. alessandra.vigano@guest.unimi.it
Lamartine Jérôme
Testoni Barbara
Merico Daniele
Alotto Daniela
Castagnoli Carlotta
Robert Amèlie
Candi Eleonora
Melino Gerry
Gidrol Xavier
Mantovani Roberto
References (60)
60 references, click to expand
  1. p63 regulates multiple signalling pathways required for ectodermal organogenesis and differentiation.
    Development. 2006 Apr;133(8):1553-63 PMID: 16524929
  2. p63 and epithelial biology.
    Exp Cell Res. 2006 Apr 1;312(6):695-706 PMID: 16406339
  3. T-bet binding to newly identified target gene promoters is cell type-independent but results in variable context-dependent functional effects.
    J Biol Chem. 2006 Apr 28;281(17):11992-2000 PMID: 16473879
  4. p63 regulates an adhesion programme and cell survival in epithelial cells.
    Nat Cell Biol. 2006 Jun;8(6):551-61 PMID: 16715076
  5. Loss of p63 leads to increased cell migration and up-regulation of genes involved in invasion and metastasis.
    Cancer Res. 2006 Aug 1;66(15):7589-97 PMID: 16885358
  6. The ferredoxin reductase gene is regulated by the p53 family and sensitizes cells to oxidative stress-induced apoptosis.
    Oncogene. 2002 Oct 17;21(47):7195-204 PMID: 12370809
  7. Complex transcriptional effects of p63 isoforms: identification of novel activation and repression domains.
    Mol Cell Biol. 2002 Dec;22(24):8659-68 PMID: 12446784
  8. REDD1, a developmentally regulated transcriptional target of p63 and p53, links p63 to regulation of reactive oxygen species.
    Mol Cell. 2002 Nov;10(5):995-1005 PMID: 12453409
  9. The Delta Np63 alpha phosphoprotein binds the p21 and 14-3-3 sigma promoters in vivo and has transcriptional repressor activity that is reduced by Hay-Wells syndrome-derived mutations.
    Mol Cell Biol. 2003 Apr;23(7):2264-76 PMID: 12640112
  10. Sarcospan-deficient mice maintain normal muscle function.
    Mol Cell Biol. 2000 Mar;20(5):1669-77 PMID: 10669744
  11. Molecular and genetic characterization of sarcospan: insights into sarcoglycan-sarcospan interactions.
    Hum Mol Genet. 2000 Aug 12;9(13):2019-27 PMID: 10942431
  12. A subset of tumor-derived mutant forms of p53 down-regulate p63 and p73 through a direct interaction with the p53 core domain.
    Mol Cell Biol. 2001 Mar;21(5):1874-87 PMID: 11238924
  13. p63 identifies keratinocyte stem cells.
    Proc Natl Acad Sci U S A. 2001 Mar 13;98(6):3156-61 PMID: 11248048
  14. Cardiac malformations, adrenal agenesis, neural crest defects and exencephaly in mice lacking Cited2, a new Tfap2 co-activator.
    Nat Genet. 2001 Dec;29(4):469-74 PMID: 11694877
  15. The p53 family member genes are involved in the Notch signal pathway.
    J Biol Chem. 2002 Jan 4;277(1):719-24 PMID: 11641404
  16. WD-repeat proteins: structure characteristics, biological function, and their involvement in human diseases.
    Cell Mol Life Sci. 2001 Dec;58(14):2085-97 PMID: 11814058
  17. p63 and p73 are required for p53-dependent apoptosis in response to DNA damage.
    Nature. 2002 Apr 4;416(6880):560-4 PMID: 11932750
  18. TAp63gamma (p51A) and dNp63alpha (p73L), two major isoforms of the p63 gene, exert opposite effects on the vascular endothelial growth factor (VEGF) gene expression.
    Oncogene. 2002 Apr 11;21(16):2455-65 PMID: 11971180
  19. A dominant-negative form of p63 is required for epidermal proliferation in zebrafish.
    Dev Cell. 2002 May;2(5):607-16 PMID: 12015968
  20. P63 gene mutations and human developmental syndromes.
    Am J Med Genet. 2002 Oct 15;112(3):284-90 PMID: 12357472
  21. Physical and functional interactions among AP-2 transcription factors, p300/CREB-binding protein, and CITED2.
    J Biol Chem. 2003 May 2;278(18):16021-9 PMID: 12586840
  22. DeltaNp63alpha and TAp63alpha regulate transcription of genes with distinct biological functions in cancer and development.
    Cancer Res. 2003 May 15;63(10):2351-7 PMID: 12750249
  23. deltaNp63alpha functions as both a positive and a negative transcriptional regulator and blocks in vitro differentiation of murine keratinocytes.
    Oncogene. 2003 Jun 5;22(23):3635-44 PMID: 12789272
  24. p73 and p63 protein stability: the way to regulate function?
    Biochem Pharmacol. 2003 Oct 15;66(8):1555-61 PMID: 14555234
  25. Gene expression of alpha-endosulfine in the rat brain: correlative changes with aging, learning and stress.
    J Neurochem. 2003 Dec;87(5):1086-100 PMID: 14622089
  26. Probing chromatin immunoprecipitates with CpG-island microarrays to identify genomic sites occupied by DNA-binding proteins.
    Methods Enzymol. 2003;371:577-96 PMID: 14712730
  27. p63 is the molecular switch for initiation of an epithelial stratification program.
    Genes Dev. 2004 Jan 15;18(2):126-31 PMID: 14729569
  28. The role of p63 in development and differentiation of the epidermis.
    J Dermatol Sci. 2004 Feb;34(1):3-9 PMID: 14757276
  29. The dynamin superfamily: universal membrane tubulation and fission molecules?
    Nat Rev Mol Cell Biol. 2004 Feb;5(2):133-47 PMID: 15040446
  30. Rat mesangial alpha-endosulfine.
    Kidney Int. 2004 May;65(5):1731-9 PMID: 15086912
  31. The common and distinct target genes of the p53 family transcription factors.
    Cell Mol Life Sci. 2004 Apr;61(7-8):822-42 PMID: 15095006
  32. Genomic approaches that aid in the identification of transcription factor target genes.
    Exp Biol Med (Maywood). 2004 Sep;229(8):705-21 PMID: 15337825
  33. p53 mutations in human immortalized epithelial cell lines.
    Carcinogenesis. 1993 May;14(5):833-9 PMID: 8504475
  34. Expression of the homeobox gene HOXC4 in keratinocytes of normal skin and epithelial skin tumors is correlated with differentiation.
    J Invest Dermatol. 1994 Sep;103(3):341-6 PMID: 7915745
  35. HOX homeobox genes exhibit spatial and temporal changes in expression during human skin development.
    J Invest Dermatol. 1998 Feb;110(2):110-5 PMID: 9457903
  36. Human alpha-endosulfine, a possible regulator of sulfonylurea-sensitive KATP channel: molecular cloning, expression and biological properties.
    Proc Natl Acad Sci U S A. 1998 Jul 7;95(14):8387-91 PMID: 9653196
  37. p63, a p53 homolog at 3q27-29, encodes multiple products with transactivating, death-inducing, and dominant-negative activities.
    Mol Cell. 1998 Sep;2(3):305-16 PMID: 9774969
  38. Functional role of p35srj, a novel p300/CBP binding protein, during transactivation by HIF-1.
    Genes Dev. 1999 Jan 1;13(1):64-75 PMID: 9887100
  39. p63 is a p53 homologue required for limb and epidermal morphogenesis.
    Nature. 1999 Apr 22;398(6729):708-13 PMID: 10227293
  40. p63 is essential for regenerative proliferation in limb, craniofacial and epithelial development.
    Nature. 1999 Apr 22;398(6729):714-8 PMID: 10227294
  41. p51/p63 Controls subunit alpha3 of the major epidermis integrin anchoring the stem cells to the niche.
    J Biol Chem. 2004 Nov 26;279(48):50069-77 PMID: 15361520
  42. IGFBP-3 is a direct target of transcriptional regulation by DeltaNp63alpha in squamous epithelium.
    Cancer Res. 2005 Mar 15;65(6):2314-20 PMID: 15781645
  43. Perp is a p63-regulated gene essential for epithelial integrity.
    Cell. 2005 Mar 25;120(6):843-56 PMID: 15797384
  44. Communicating with Hedgehogs.
    Nat Rev Mol Cell Biol. 2005 Apr;6(4):306-17 PMID: 15803137
  45. Chromatin immunoprecipitation (ChIP) on chip experiments uncover a widespread distribution of NF-Y binding CCAAT sites outside of core promoters.
    J Biol Chem. 2005 Apr 8;280(14):13606-15 PMID: 15647281
  46. Devising transcriptional regulatory networks operating during the cell cycle and differentiation using ChIP-on-chip.
    Chromosome Res. 2005;13(3):275-88 PMID: 15868421
  47. CpG Island microarray probe sequences derived from a physical library are representative of CpG Islands annotated on the human genome.
    Nucleic Acids Res. 2005;33(9):2952-61 PMID: 15911630
  48. Differential recognition of response elements determines target gene specificity for p53 and p63.
    Mol Cell Biol. 2005 Jul;25(14):6077-89 PMID: 15988020
  49. TAp63alpha induces apoptosis by activating signaling via death receptors and mitochondria.
    EMBO J. 2005 Jul 6;24(13):2458-71 PMID: 15944736
  50. Identification of pigment epithelium-derived factor as a direct target of the p53 family member genes.
    Oncogene. 2005 Jul 28;24(32):5131-6 PMID: 15856012
  51. The sarcoglycan-sarcospan complex localization in mouse retina is independent from dystrophins.
    Neurosci Res. 2005 Sep;53(1):25-33 PMID: 15993965
  52. Regulatory role of dynamin-2 in VEGFR-2/KDR-mediated endothelial signaling.
    FASEB J. 2005 Oct;19(12):1692-4 PMID: 16049137
  53. Hypoxia up-regulates expression of Eph receptors and ephrins in mouse skin.
    FASEB J. 2005 Oct;19(12):1689-91 PMID: 16081502
  54. Regulation of the cyclin-dependent kinase inhibitor p57Kip2 expression by p63.
    Cell Cycle. 2005 Nov;4(11):1625-31 PMID: 16258268
  55. A novel response element confers p63- and p73-specific activation of the WNT4 promoter.
    Biochem Biophys Res Commun. 2006 Jan 27;339(4):1120-8 PMID: 16343436
  56. TAp63alpha induces AP-2gamma as an early event in epidermal morphogenesis.
    Dev Biol. 2006 Jan 1;289(1):253-61 PMID: 16324689
  57. AP-2alpha: a regulator of EGF receptor signaling and proliferation in skin epidermis.
    J Cell Biol. 2006 Jan 30;172(3):409-21 PMID: 16449191
  58. p53 ubiquitination: Mdm2 and beyond.
    Mol Cell. 2006 Feb 3;21(3):307-15 PMID: 16455486
  59. Genetic elimination of Suppressor of fused reveals an essential repressor function in the mammalian Hedgehog signaling pathway.
    Dev Cell. 2006 Feb;10(2):187-97 PMID: 16459298
  60. Cross-regulation between Notch and p63 in keratinocyte commitment to differentiation.
    Genes Dev. 2006 Apr 15;20(8):1028-42 PMID: 16618808
Article Info
Journal
The EMBO journal
Abbr.
EMBO J
ISSN
0261-4189
DOI
10.1038/sj.emboj.7601375
Email
alessandra.vigano@guest.unimi.it
Published
2006-11-01
Epub
2006-00-12
Pages
5105-16
Language
English
Region
England
NLM ID
8208664
PMCID
PMC1630419
Subset
IM
Grants
Telethon · GGP02379 · Italy
Medical Research Council · MC_U132670600 · United Kingdom
External Links
PubMed source DOI: 10.1038/sj.emboj.7601375 PMC Full Text View journal details
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