Binding to Nonmethylated CpG DNA Is Essential for Target Recognition, Transactivation, and Myeloid Transformation by an MLL Oncoprotein

ABSTRACT The MLL gene is a frequent target for leukemia-associated chromosomal translocations that generate dominant-acting chimeric oncoproteins. These invariably contain the amino-terminal 1,400 residues of MLL fused with one of a variety of over 30 distinct nuclear or cytoplasmic partner proteins. Despite the consistent inclusion of the MLL amino-terminal region in leukemia oncoproteins, little is known regarding its molecular contributions to MLL-dependent oncogenesis. Using high-resolution mutagenesis, we identified three MLL domains that are essential for in vitro myeloid transformation via mechanisms that do not compromise subnuclear localization. These include the CXXC/Basic domain and two novel domains of unknown function. Point mutations in the CXXC domain that eliminate myeloid transformation by an MLL fusion protein also abolished recognition and binding of nonmethylated CpG DNA sites in vitro and transactivation in vivo. Our results define a critical role for the CXXC DNA binding domain in MLL-associated oncogenesis, most likely via epigenetic recognition of CpG DNA sites within the regulatory elements of target genes.

[1]  M. Cleary,et al.  Transformation of myeloid progenitors by MLL oncoproteins is dependent on Hoxa7 and Hoxa9. , 2003, Genes & development.

[2]  M. Cleary,et al.  Dimerization contributes to oncogenic activation of MLL chimeras in acute leukemias. , 2003, Cancer cell.

[3]  M. García-Cuéllar,et al.  Transcriptional activation is a key function encoded by MLL fusion partners , 2003, Leukemia.

[4]  Chi Wai So,et al.  Common mechanism for oncogenic activation of MLL by forkhead family proteins. , 2003, Blood.

[5]  Toshiki Mori,et al.  ALL-1 is a histone methyltransferase that assembles a supercomplex of proteins involved in transcriptional regulation. , 2002, Molecular cell.

[6]  Thomas A Milne,et al.  MLL targets SET domain methyltransferase activity to Hox gene promoters. , 2002, Molecular cell.

[7]  M. Cleary,et al.  MLL-AFX Requires the Transcriptional Effector Domains of AFX To Transform Myeloid Progenitors and Transdominantly Interfere with Forkhead Protein Function , 2002, Molecular and Cellular Biology.

[8]  E. Green,et al.  MLL5, a homolog of Drosophila trithorax located within a segment of chromosome band 7q22 implicated in myeloid leukemia , 2002, Oncogene.

[9]  M. Cleary,et al.  The AF10 leucine zipper is required for leukemic transformation of myeloid progenitors by MLL-AF10. , 2002, Blood.

[10]  J. Downing,et al.  Classification, subtype discovery, and prediction of outcome in pediatric acute lymphoblastic leukemia by gene expression profiling. , 2002, Cancer cell.

[11]  M. Birke,et al.  The MT domain of the proto-oncoprotein MLL binds to CpG-containing DNA and discriminates against methylation. , 2002, Nucleic acids research.

[12]  Gérard Roizès,et al.  MLL3, a new human member of the TRX/MLL gene family, maps to 7q36, a chromosome region frequently deleted in myeloid leukaemia. , 2002, Gene.

[13]  R. Humphries,et al.  Differential expression of Hox, Meis1, and Pbx1 genes in primitive cells throughout murine hematopoietic ontogeny. , 2002, Experimental hematology.

[14]  E. Lander,et al.  MLL translocations specify a distinct gene expression profile that distinguishes a unique leukemia , 2002, Nature Genetics.

[15]  A. Bird.  DNA methylation patterns and epigenetic memory. , 2002, Genes & development.

[16]  K. Voo,et al.  Identification and Characterization of the DNA Binding Domain of CpG-binding Protein* , 2001, The Journal of Biological Chemistry.

[17]  M. Cleary,et al.  Molecular mechanisms of leukemogenesis mediated by MLL fusion proteins , 2001, Oncogene.

[18]  C. Lavau,et al.  The Elongation Domain of ELL Is Dispensable but Its ELL-Associated Factor 1 Interaction Domain Is Essential for MLL-ELL-Induced Leukemogenesis , 2001, Molecular and Cellular Biology.

[19]  M. Owen,et al.  Truncation of the MLL gene in exon 5 by gene targeting leads to early preimplantation lethality of homozygous embryos , 2001, Genesis.

[20]  C. Croce,et al.  Upregulation of Meis1 and HoxA9 in acute lymphocytic leukemias with the t(4 : 11) abnormality , 2001, Oncogene.

[21]  A. Shilatifard,et al.  A carboxy-terminal domain of ELL is required and sufficient for immortalization of myeloid progenitors by MLL-ELL. , 2000, Blood.

[22]  Changchun Du,et al.  Chromatin‐related properties of CBP fused to MLL generate a myelodysplastic‐like syndrome that evolves into myeloid leukemia , 2000, The EMBO journal.

[23]  M. Nakao,et al.  Tsutomu by Methyl-CpG Binding Protein MBD 1 Mechanism of Transcriptional Regulation , 2000 .

[24]  S. Chin,et al.  MLL2, the second human homolog of the Drosophila trithorax gene, maps to 19q13.1 and is amplified in solid tumor cell lines , 1999, Oncogene.

[25]  C. Allis,et al.  Methylation of histone H3 at lysine 4 is highly conserved and correlates with transcriptionally active nuclei in Tetrahymena. , 1999, Proceedings of the National Academy of Sciences of the United States of America.

[26]  S. Schreiner,et al.  The leukemogenic fusion of MLL with ENL creates a novel transcriptional transactivator , 1999, Leukemia.

[27]  M. Diaz,et al.  MLL2: A new mammalian member of the trx/MLL family of genes. , 1999, Genomics.

[28]  R. D. Hanson,et al.  MLL, a mammalian trithorax-group gene, functions as a transcriptional maintenance factor in morphogenesis. , 1998, Proceedings of the National Academy of Sciences of the United States of America.

[29]  T. Komori,et al.  Growth disturbance in fetal liver hematopoiesis of Mll-mutant mice. , 1998, Blood.

[30]  C. Caldas,et al.  Isolation and characterization of a Pufferfish MLL (Mixed lineage leukemia)-like gene (fMll) reveals evolutionary conservation in vertebrate genes related to Drosophila trithorax , 1998, Oncogene.

[31]  Michael L. Cleary,et al.  The Oncogenic Capacity of HRX-ENL Requires the Transcriptional Transactivation Activity of ENL and the DNA Binding Motifs of HRX , 1998, Molecular and Cellular Biology.

[32]  Jeffrey R. Huth,et al.  The solution structure of an HMG-I(Y)–DNA complex defines a new architectural minor groove binding motif , 1997, Nature Structural Biology.

[33]  C. Croce,et al.  Structure and expression pattern of human ALR, a novel gene with strong homology to ALL-1 involved in acute leukemia and to Drosophila trithorax , 1997, Oncogene.

[34]  M. Cleary,et al.  Immortalization and leukemic transformation of a myelomonocytic precursor by retrovirally transduced HRX–ENL , 1997, The EMBO journal.

[35]  C. Dang,et al.  Nuclear punctate distribution of ALL-1 is conferred by distinct elements at the N terminus of the protein. , 1997, Proceedings of the National Academy of Sciences of the United States of America.

[36]  T. Magnuson,et al.  Murine Polycomb- and trithorax-group genes regulate homeotic pathways and beyond. , 1997, Trends in genetics : TIG.

[37]  S. Korsmeyer,et al.  Altered Hox expression and segmental identity in Mll-mutant mice , 1995, Nature.

[38]  G. Sauvageau,et al.  Stage- and lineage-specific expression of the HOXA10 homeobox gene in normal and leukemic hematopoietic cells. , 1995, Experimental hematology.

[39]  G. Sauvageau,et al.  Differential expression of homeobox genes in functionally distinct CD34+ subpopulations of human bone marrow cells. , 1994, Proceedings of the National Academy of Sciences of the United States of America.

[40]  S. Korsmeyer,et al.  Acute mixed-lineage leukemia t(4;11)(q21;q23) generates an MLL-AF4 fusion product. , 1993, Proceedings of the National Academy of Sciences of the United States of America.

[41]  H. Alder,et al.  The t(4;11) chromosome translocation of human acute leukemias fuses the ALL-1 gene, related to Drosophila trithorax, to the AF-4 gene , 1992, Cell.

[42]  Michael L. Cleary,et al.  Involvement of a homolog of Drosophila trithorax by 11q23 chromosomal translocations in acute leukemias , 1992, Cell.

[43]  G. Evans,et al.  A trithorax–like gene is interrupted by chromosome 11q23 translocations in acute leukaemias , 1992, Nature Genetics.