E-box-independent regulation of transcription and differentiation by MYC

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MYC proto-oncogene is a key player in cell homeostasis that is commonly deregulated in human carcinogenesis1. MYC can either activate or repress target genes by forming a complex with MAX (ref. 2). MYC also exerts MAX-independent functions that are not yet fully characterized3. Cells possess an intrinsic pathway that can abrogate MYC–MAX dimerization and E-box interaction, by inducing phosphorylation of MYC in a PAK2-dependent manner at three residues located in its helix–loop–helix domain4. Here we show that these carboxy-terminal phosphorylation events switch MYC from an oncogenic to a tumour-suppressive function. In undifferentiated cells, MYC–MAX is targeted to the promoters of retinoic-acid-responsive genes by its direct interaction with the retinoic acid receptor-α (RARα). MYC–MAX cooperates with RARα to repress genes required for differentiation, in an E-box-independent manner. Conversely, on C-terminal phosphorylation of MYC during differentiation, the complex switches from a repressive to an activating function, by releasing MAX and recruiting transcriptional co-activators. Phospho-MYC synergizes with retinoic acid to eliminate circulating leukaemic cells and to decrease the level of tumour invasion. Our results identify an E-box-independent mechanism for transcriptional regulation by MYC that unveils previously unknown functions for MYC in differentiation. These may be exploited to develop alternative targeted therapies.

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Figure 1: MYC regulates RA-target genes in a phosphorylation-dependent manner.
Figure 2: MYC regulates RARE-containing genes by binding to RARα.
Figure 3: MYC recruits co-regulators to RARα-target genes and is phosphorylated by activated PAK2 during cell differentiation.
Figure 4: Phosphorylation of MYC by PAK2 is required for proper RA-target gene activation and cell differentiation.
Figure 5: Phospho-MYC potentates RA therapy in leukaemia, both in vitro and in vivo.


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We thank T. Berg and C. Rochette-Egly for reagents, P. Muñoz-Cánoves, S. de la Luna and G. Gil for discussions, S. Colomer-Lahiguera and J. Douet for graphical assistance, V. Valero for technical support and V. A. Raker for helpful discussions about the manuscript. This work was supported by grants from the ‘Fondo de Investigación Sanitario’ (FIS, Spanish Ministerio de Sanidad) to S.A.B.; and from the Spanish ‘Ministerio de Educación y Ciencia’, AGAUR and ‘Fundació La Marató’ to L.D.C. I.U. has been supported by an FPI fellowship of the Spanish Ministerio de Ciencia y Educación, M.B. by a DFG and a Ramón y Cajal fellowship, S.T. by a ‘La Caixa’ fellowship and S.D. by a PFIS fellowship.

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I.U. and M.B. carried out reporter assays, co-immunoprecipitations and ChIPs; I.U. and A.G. conducted cellular infections, flow cytometry analyses and gene expression experiments as well as differentiation studies with assistance from J.F.N; I.U., M.B., S.T. and S.D. carried out recombinant protein expression, pulldown experiments and phosphorylation assays; B.K. carried out Flag-affinity purification and generated HEK293T MYC–ER stable clones; J.M-C. carried out the experimental work on mice, assisted by I.U. and A.G.; G.R. carried out all bioinformatic analyses; L.D.C., S.A.B. and I.U. supervised the project and designed the experiments; I.U., M.B., S.A.B. and L.D.C. wrote the manuscript.

Correspondence to Salvador Aznar Benitah or Luciano Di Croce.

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Uribesalgo, I., Buschbeck, M., Gutiérrez, A. et al. E-box-independent regulation of transcription and differentiation by MYC. Nat Cell Biol 13, 1443–1449 (2011) doi:10.1038/ncb2355

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