The controls that enable melanoblasts and melanoma cells to proliferate are likely to be related, but so far no key regulator of cell cycle progression specific to the melanocyte lineage has been identified. The microphthalmia-associated transcription factor Mitf has a crucial but poorly defined role in melanoblast and melanocyte survival and in differentiation1. Here we show that Mitf can act as a novel anti-proliferative transcription factor able to induce a G1 cell-cycle arrest that is dependent on Mitf-mediated activation of the p21Cip1 (CDKN1A) cyclin-dependent kinase inhibitor gene. Moreover, cooperation between Mitf and the retinoblastoma protein Rb1 potentiates the ability of Mitf to activate transcription. The results indicate that Mitf-mediated activation of p21Cip1 expression and consequent hypophosphorylation of Rb1 will contribute to cell cycle exit and activation of the differentiation programme. The mutation of genes associated with melanoma, such as INK4a or BRAF that would affect either Mitf cooperation with Rb1 or Mitf stability respectively, would impair Mitf-mediated cell cycle control.
Access optionsAccess options
Subscribe to Journal
Get full journal access for 1 year
only $3.90 per issue
All prices are NET prices.
VAT will be added later in the checkout.
Rent or Buy article
Get time limited or full article access on ReadCube.
All prices are NET prices.
Vance, K. W. & Goding, C. R. The transcription network regulating melanocyte development and melanoma. Pigment Cell Res. 17, 318–325 (2004)
Bertolotto, C. et al. Microphthalmia gene product as a signal transducer in cAMP-induced differentiation of melanocytes. J. Cell Biol. 142, 827–835 (1998)
Littlewood, T. D., Hancock, D. C., Danielian, P. S., Parker, M. G. & Evan, G. I. A modified oestrogen receptor ligand-binding domain as an improved switch for the regulation of heterologous proteins. Nucleic Acids Res. 23, 1686–1690 (1995)
Quelle, D. E. et al. Cloning and characterization of murine p16INK4a and p15INK4b genes. Oncogene 11, 635–645 (1995)
Quelle, D. E., Zindy, F., Ashmun, R. A. & Sherr, C. J. Alternative reading frames of the INK4a tumor suppressor gene encode two unrelated proteins capable of inducing cell cycle arrest. Cell 83, 993–1000 (1995)
Halaban, R., Cheng, E., Zhang, Y., Mandigo, C. E. & Miglarese, M. R. Release of cell cycle constraints in mouse melanocytes by overexpressed mutant E2F1E132, but not by deletion of p16INK4a or p21WAF1/CIP1. Oncogene 16, 2489–2501 (1998)
McGill, G. G. et al. Bcl2 regulation by the melanocyte master regulator Mitf modulates lineage survival and melanoma cell viability. Cell 109, 707–718 (2002)
Aksan, I. & Goding, C. R. Targeting the microphthalmia basic helix-loop-helix-leucine zipper transcription factor to a subset of E-box elements in vitro and in vivo. Mol. Cell. Biol. 18, 6930–6938 (1998)
Bentley, N. J., Eisen, T. & Goding, C. R. Melanocyte-specific expression of the human tyrosinase promoter: activation by the microphthalmia gene product and role of the initiator. Mol. Cell. Biol. 14, 7996–8006 (1994)
Allan, L. A., Duhig, T., Read, M. & Fried, M. The p21(WAF1/CIP1) promoter is methylated in Rat-1 cells: stable restoration of p53-dependent p21(WAF1/CIP1) expression after transfection of a genomic clone containing the p21(WAF1/CIP1) gene. Mol. Cell. Biol. 20, 1291–1298 (2000)
Deng, C., Zhang, P., Harper, J. W., Elledge, S. J. & Leder, P. Mice lacking p21CIP1/WAF1 undergo normal development, but are defective in G1 checkpoint control. Cell 82, 675–684 (1995)
Yu, B. D. et al. Distinct and nonoverlapping roles for pRB and cyclin D:cyclin-dependent kinases 4/6 activity in melanocyte survival. Proc. Natl Acad. Sci. USA 100, 14881–14886 (2003)
Lerner, A. B. et al. A mouse model for vitiligo. J. Invest. Dermatol. 87, 299–304 (1986)
Yavuzer, U. et al. The microphthalmia gene product interacts with the retinoblastoma protein in vitro and is a target for deregulation of melanocyte-specific transcription. Oncogene 10, 123–134 (1995)
Chew, Y. P., Ellis, M., Wilkie, S. & Mittnacht, S. pRB phosphorylation mutants reveal role of pRB in regulating S phase completion by a mechanism independent of E2F. Oncogene 17, 2177–2186 (1998)
Takahashi, Y., Rayman, J. B. & Dynlacht, B. D. Analysis of promoter binding by the E2F and pRB families in vivo: distinct E2F proteins mediate activation and repression. Genes Dev. 14, 804–816 (2000)
Umek, R. M., Friedman, A. D. & McKnight, S. L. CCAAT-enhancer binding protein: a component of a differentiation switch. Science 251, 288–292 (1991)
Okura, M., Maeda, H., Nishikawa, S. & Mizoguchi, M. Effects of monoclonal anti-c-kit antibody (ACK2) on melanocytes in newborn mice. J. Invest. Dermatol. 105, 322–328 (1995)
Hornyak, T. J., Hayes, D. J., Chiu, L. Y. & Ziff, E. B. Transcription factors in melanocyte development: distinct roles for Pax-3 and Mitf. Mech. Dev. 101, 47–59 (2001)
Konyukhov, B. V. & Sazhina, M. V. Interaction of the genes of ocular retardation and microphthalmia in mice. Folia (Praha) 12, 116–123 (1966)
Davies, H. et al. Mutations of the BRAF gene in human cancer. Nature 417, 949–954 (2002)
Jiang, H. et al. The melanoma differentiation-associated gene mda-6, which encodes the cyclin-dependent kinase inhibitor p21, is differentially expressed during growth, differentiation and progression in human melanoma cells. Oncogene 10, 1855–1864 (1995)
Hodgkinson, C. A. et al. Mutations at the mouse microphthalmia locus are associated with defects in a gene encoding a novel basic-helix–loop–helix-zipper protein. Cell 74, 395–404 (1993)
Carreira, S., Liu, B. & Goding, C. R. The gene encoding the T-box transcription factor Tbx2 is a target for the microphthalmia-associated transcription factor in melanocytes. J. Biol. Chem. 275, 21920–21927 (2000)
Prince, S., Carreira, S., Vance, K. W., Abrahams, A. & Goding, C. R. Tbx2 directly represses the expression of the p21(WAF1) cyclin-dependent kinase inhibitor. Cancer Res. 64, 1669–1674 (2004)
Jacobs, J. J. et al. Senescence bypass screen identifies TBX2, which represses cdkn2a (p19ARF) and is amplified in a subset of human breast cancers. Nature Genet. 26, 291–299 (2000)
Vigo, E. et al. CDC25A phosphatase is a target of E2F and is required for efficient E2F-induced S phase. Mol. Cell. Biol. 19, 6379–6395 (1999)
Carreira, S., Dexter, T. J., Yavuzer, U., Easty, D. J. & Goding, C. R. Brachyury-related transcription factor Tbx2 and repression of the melanocyte-specific TRP-1 promoter. Mol. Cell. Biol. 18, 5099–5108 (1998)
Hirst, K., Fisher, F., McAndrew, P. C. & Goding, C. R. The transcription factor, the Cdk, its cyclin and their regulator: directing the transcriptional response to a nutritional signal. EMBO J. 13, 5410–5420 (1994)
Galibert, M. D., Carreira, S. & Goding, C. R. The Usf-1 transcription factor is a novel target for the stress-responsive p38 kinase and mediates UV-induced tyrosinase expression. EMBO J. 20, 5022–5031 (2001)
We thank M. Serrano for the WT and p21-null MEFs; S. Mittnacht for the Rb1 expression vectors and the C33a cells; B. Vogelstein for the p21Cip1 promoter; K. Helin for the HA.ER-expression vector; D. Stillman for yeast reporter strain DY1641; C. Wellbrock, R. Marais, R. Ballotti and C. Bertolotto for communication of unpublished results; and H. Arnheiter for discussions. This work was supported by Marie Curie Cancer Care, the Association for International Cancer Research and a European Union Marie Curie fellowship to M.-D.G.
The authors declare that they have no competing financial interests.
About this article
Hydrogen Sulfide Inhibits Formaldehyde-Induced Senescence in HT-22 Cells via Upregulation of Leptin Signaling
NeuroMolecular Medicine (2019)
Quantitative Proteomics Reveals Changes in Vero Cells in Response to Porcine Epidemic Diarrhea Virus
Journal of Proteome Research (2019)
Yin Yang 1 Orchestrates a Metabolic Program Required for Both Neural Crest Development and Melanoma Formation
Cell Stem Cell (2019)
BAF60A mediates interactions between the microphthalmia‐associated transcription factor and the BRG1‐containing SWI/SNF complex during melanocyte differentiation
Journal of Cellular Physiology (2019)