Transcriptional repression of miR-34 family contributes to p63-mediated cell cycle progression in epidermal cells.

p63, a p53 family member, is highly expressed in the basal proliferative compartment of the epidermis and its expression has been correlated with the growth ability and regenerative capacity of keratinocytes. In this study we report a mechanism through which p63 maintains cell cycle progression by directly repressing miR-34a and miR-34c. In the absence of p63, increased levels of miR-34a and miR-34c were observed in primary keratinocytes and in embryonic skin, with concomitant G1-phase arrest and inhibition of the cell cycle regulators cyclin D1 and cyclin-dependent kinase 4 (Cdk4). p63 directly bound to p53-consensus sites in both miR-34a and miR-34c regulatory regions and inhibited their activity. Concomitant downregulation of miR-34a and miR-34c substantially restored cell cycle progression and expression of cyclin D1 and Cdk4. Our data indicate that specific miR-34 family members have a significant role downstream of p63 in controlling epidermal cell proliferation.

[1]  J. Pietenpol,et al.  Transcriptional Programs Regulated by p63 in Normal Epithelium and Tumors , 2007, Cell cycle.

[2]  E. Flores,et al.  Rescue of key features of the p63‐null epithelial phenotype by inactivation of Ink4a and Arf , 2009, The EMBO journal.

[3]  D. Bartel.  MicroRNAs: Target Recognition and Regulatory Functions , 2009, Cell.

[4]  S. Elledge,et al.  The p21 Cdk-interacting protein Cip1 is a potent inhibitor of G1 cyclin-dependent kinases , 1993, Cell.

[5]  F. McKeon,et al.  p63 Is Essential for the Proliferative Potential of Stem Cells in Stratified Epithelia , 2007, Cell.

[6]  A. Mills,et al.  p63 is the molecular switch for initiation of an epithelial stratification program. , 2004, Genes & development.

[7]  Moshe Oren,et al.  Transcriptional activation of miR-34a contributes to p53-mediated apoptosis. , 2007, Molecular cell.

[8]  K. Kinzler,et al.  p21 is necessary for the p53-mediated G1 arrest in human cancer cells. , 1995, Cancer research.

[9]  Naoto Tsuchiya,et al.  Tumor-suppressive miR-34a induces senescence-like growth arrest through modulation of the E2F pathway in human colon cancer cells , 2007, Proceedings of the National Academy of Sciences.

[10]  A. Yang,et al.  p63, a p53 homolog at 3q27-29, encodes multiple products with transactivating, death-inducing, and dominant-negative activities. , 1998, Molecular cell.

[11]  Rebecca A. Ihrie,et al.  Perp Is a p63-Regulated Gene Essential for Epithelial Integrity , 2005, Cell.

[12]  J. Pietenpol,et al.  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. , 2003, Molecular and cellular biology.

[13]  A. Yang,et al.  A C-Terminal Inhibitory Domain Controls the Activity of p63 by an Intramolecular Mechanism , 2002, Molecular and Cellular Biology.

[14]  D. Ponzin,et al.  p63 identifies keratinocyte stem cells , 2001, Proceedings of the National Academy of Sciences of the United States of America.

[15]  J. Pietenpol,et al.  The ΔNp63α Phosphoprotein Binds the p21 and 14-3-3σ Promoters In Vivo and Has Transcriptional Repressor Activity That Is Reduced by Hay-Wells Syndrome-Derived Mutations , 2003, Molecular and Cellular Biology.

[16]  Wei Wang,et al.  MicroRNA-34b and MicroRNA-34c are targets of p53 and cooperate in control of cell proliferation and adhesion-independent growth. , 2007, Cancer research.

[17]  D. Merico,et al.  New p63 targets in keratinocytes identified by a genome‐wide approach , 2006, The EMBO journal.

[18]  Michael A. Beer,et al.  Transactivation of miR-34a by p53 broadly influences gene expression and promotes apoptosis. , 2007, Molecular cell.

[19]  Elaine Fuchs,et al.  A skin microRNA promotes differentiation by repressing ‘stemness’ , 2008, Nature.

[20]  R. Knight,et al.  miR-203 represses ‘stemness’ by repressing ΔNp63 , 2008, Cell Death and Differentiation.

[21]  Jian Wang,et al.  Cross-regulation between Notch and p63 in keratinocyte commitment to differentiation. , 2006, Genes & development.

[22]  M. Kretz,et al.  p63 regulates proliferation and differentiation of developmentally mature keratinocytes. , 2006, Genes & development.

[23]  J. Trent,et al.  WAF1, a potential mediator of p53 tumor suppression , 1993, Cell.

[24]  James Brugarolas,et al.  Radiation-induced cell cycle arrest compromised by p21 deficiency , 1995, Nature.

[25]  Jason S. Carroll,et al.  p63 regulates an adhesion programme and cell survival in epithelial cells , 2006, Nature Cell Biology.

[26]  Chee-Onn Leong,et al.  Tumor-specific p73 up-regulation mediates p63 dependence in squamous cell carcinoma. , 2006, Cancer research.

[27]  R. Buller,et al.  The Relationship of Molecular Markers of p53 Function and Angiogenesis to Prognosis of Stage I Epithelial Ovarian Cancer , 2005, Clinical Cancer Research.

[28]  V. Tarasov,et al.  Differential Regulation of microRNAs by p53 Revealed by Massively Parallel Sequencing: miR-34a is a p53 Target That Induces Apoptosis and G1-arrest , 2007, Cell cycle.

[29]  Ying Feng,et al.  Supplemental Data P53-mediated Activation of Mirna34 Candidate Tumor-suppressor Genes , 2022 .

[30]  F. Dietrich,et al.  Morphogenesis in skin is governed by discrete sets of differentially expressed microRNAs , 2006, Nature Genetics.

[31]  Eduardo Sontag,et al.  Transcriptional control of human p53-regulated genes , 2008, Nature Reviews Molecular Cell Biology.

[32]  Hiroshi I. Suzuki,et al.  Modulation of microRNA processing by p53 , 2009, Nature.

[33]  M. Dohn,et al.  p63alpha and DeltaNp63alpha can induce cell cycle arrest and apoptosis and differentially regulate p53 target genes. , 2001, Oncogene.

[34]  C. Missero,et al.  Requirement of the forkhead gene Foxe1, a target of sonic hedgehog signaling, in hair follicle morphogenesis. , 2004, Human molecular genetics.

[35]  M. Zannini,et al.  An Autoregulatory Loop Directs the Tissue-Specific Expression of p63 through a Long-Range Evolutionarily Conserved Enhancer , 2006, Molecular and Cellular Biology.

[36]  Christopher P. Crum,et al.  p63 is essential for regenerative proliferation in limb, craniofacial and epithelial development , 1999, Nature.

[37]  L. Donehower,et al.  Mice deficient for p53 are developmentally normal but susceptible to spontaneous tumours , 1992, Nature.

[38]  L. Lim,et al.  A microRNA component of the p53 tumour suppressor network , 2007, Nature.

[39]  D. di Bernardo,et al.  Direct targets of the TRP63 transcription factor revealed by a combination of gene expression profiling and reverse engineering. , 2008, Genome research.

[40]  Yi Tie,et al.  Downregulation of CCND1 and CDK6 by miR‐34a induces cell cycle arrest , 2008, FEBS letters.

[41]  E. Stupka,et al.  Tprg, a gene predominantly expressed in skin, is a direct target of the transcription factor p63. , 2008, The Journal of investigative dermatology.

[42]  Y. Nagakawa,et al.  p63-specific activation of the BPAG-1e promoter. , 2005, The Journal of investigative dermatology.

[43]  H. Vogel,et al.  p63 is a p53 homologue required for limb and epidermal morphogenesis , 1999, Nature.

[44]  C. Sander,et al.  DGCR8-dependent microRNA biogenesis is essential for skin development , 2009, Proceedings of the National Academy of Sciences.

[45]  S. Lowe,et al.  p63 deficiency activates a program of cellular senescence and leads to accelerated aging. , 2005, Genes & development.

[46]  A. Yang,et al.  REDD1, a developmentally regulated transcriptional target of p63 and p53, links p63 to regulation of reactive oxygen species. , 2002, Molecular cell.

[47]  Xinbin Chen,et al.  p63α and ΔNp63α can induce cell cycle arrest and apoptosis and differentially regulate p53 target genes , 2001, Oncogene.

[48]  R. Yi,et al.  MicroRNA-mediated control in the skin , 2010, Cell Death and Differentiation.

[49]  G. Hannon,et al.  The miRNA-Processing Enzyme Dicer Is Essential for the Morphogenesis and Maintenance of Hair Follicles , 2006, Current Biology.

[50]  Kevin Struhl,et al.  Relationships between p63 binding, DNA sequence, transcription activity, and biological function in human cells. , 2006, Molecular cell.

[51]  F. McKeon,et al.  Association of p63 with proliferative potential in normal and neoplastic human keratinocytes. , 1999, The Journal of investigative dermatology.