A DNMT3A Mutation Common in AML Exhibits Dominant-negative Effects in Murine ES Cells

Overview

Journal Blood

Publisher Elsevier

Specialty Hematology

Date 2013 Oct 30

PMID 24167195

Citations 117

Authors

Soo Jin Kim

Hongbo Zhao

Swanand Hardikar

Anup Kumar Singh

Margaret A Goodell

Taiping Chen

Affiliations

Soon will be listed here.

Abstract

Somatic heterozygous mutations of the DNA methyltransferase gene DNMT3A occur frequently in acute myeloid leukemia and other hematological malignancies, with the majority (∼60%) of mutations affecting a single amino acid, Arg882 (R882), in the catalytic domain. Although the mutations impair DNMT3A catalytic activity in vitro, their effects on DNA methylation in cells have not been explored. Here, we show that exogenously expressed mouse Dnmt3a proteins harboring the corresponding R878 mutations largely fail to mediate DNA methylation in murine embryonic stem (ES) cells but are capable of interacting with wild-type Dnmt3a and Dnmt3b. Coexpression of the Dnmt3a R878H (histidine) mutant protein results in inhibition of the ability of wild-type Dnmt3a and Dnmt3b to methylate DNA in murine ES cells. Furthermore, expression of Dnmt3a R878H in ES cells containing endogenous Dnmt3a or Dnmt3b induces hypomethylation. These results suggest that the DNMT3A R882 mutations, in addition to being hypomorphic, have dominant-negative effects.

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References

Yamashita Y, Yuan J, Suetake I, Suzuki H, Ishikawa Y, Choi Y . Array-based genomic resequencing of human leukemia. Oncogene. 2010; 29(25):3723-31. DOI: 10.1038/onc.2010.117. View

Kim J, Lee S, Li L, Park H, Park J, Lee K . High cleavage efficiency of a 2A peptide derived from porcine teschovirus-1 in human cell lines, zebrafish and mice. PLoS One. 2011; 6(4):e18556. PMC: 3084703. DOI: 10.1371/journal.pone.0018556. View

Grossmann V, Haferlach C, Weissmann S, Roller A, Schindela S, Poetzinger F . The molecular profile of adult T-cell acute lymphoblastic leukemia: mutations in RUNX1 and DNMT3A are associated with poor prognosis in T-ALL. Genes Chromosomes Cancer. 2013; 52(4):410-22. DOI: 10.1002/gcc.22039. View

Challen G, Sun D, Jeong M, Luo M, Jelinek J, Berg J . Dnmt3a is essential for hematopoietic stem cell differentiation. Nat Genet. 2011; 44(1):23-31. PMC: 3637952. DOI: 10.1038/ng.1009. View

Stegelmann F, Bullinger L, Schlenk R, Paschka P, Griesshammer M, Blersch C . DNMT3A mutations in myeloproliferative neoplasms. Leukemia. 2011; 25(7):1217-9. DOI: 10.1038/leu.2011.77. View

Okano M, BELL D, Haber D, Li E . DNA methyltransferases Dnmt3a and Dnmt3b are essential for de novo methylation and mammalian development. Cell. 1999; 99(3):247-57. DOI: 10.1016/s0092-8674(00)81656-6. View

Gaidzik V, Schlenk R, Paschka P, Stolzle A, Spath D, Kuendgen A . Clinical impact of DNMT3A mutations in younger adult patients with acute myeloid leukemia: results of the AML Study Group (AMLSG). Blood. 2013; 121(23):4769-77. DOI: 10.1182/blood-2012-10-461624. View

Li J, Pu M, Hirasawa R, Li B, Huang Y, Zeng R . Synergistic function of DNA methyltransferases Dnmt3a and Dnmt3b in the methylation of Oct4 and Nanog. Mol Cell Biol. 2007; 27(24):8748-59. PMC: 2169413. DOI: 10.1128/MCB.01380-07. View

Abdel-Wahab O, Pardanani A, Rampal R, Lasho T, Levine R, Tefferi A . DNMT3A mutational analysis in primary myelofibrosis, chronic myelomonocytic leukemia and advanced phases of myeloproliferative neoplasms. Leukemia. 2011; 25(7):1219-20. PMC: 4640464. DOI: 10.1038/leu.2011.82. View

10.

Walter M, Ding L, Shen D, Shao J, Grillot M, McLellan M . Recurrent DNMT3A mutations in patients with myelodysplastic syndromes. Leukemia. 2011; 25(7):1153-8. PMC: 3202965. DOI: 10.1038/leu.2011.44. View

11.

Chedin F, Lieber M, Hsieh C . The DNA methyltransferase-like protein DNMT3L stimulates de novo methylation by Dnmt3a. Proc Natl Acad Sci U S A. 2002; 99(26):16916-21. PMC: 139244. DOI: 10.1073/pnas.262443999. View

12.

Chen T, Ueda Y, Dodge J, Wang Z, Li E . Establishment and maintenance of genomic methylation patterns in mouse embryonic stem cells by Dnmt3a and Dnmt3b. Mol Cell Biol. 2003; 23(16):5594-605. PMC: 166327. DOI: 10.1128/MCB.23.16.5594-5605.2003. View

13.

Jurkowska R, Rajavelu A, Anspach N, Urbanke C, Jankevicius G, Ragozin S . Oligomerization and binding of the Dnmt3a DNA methyltransferase to parallel DNA molecules: heterochromatic localization and role of Dnmt3L. J Biol Chem. 2011; 286(27):24200-7. PMC: 3129201. DOI: 10.1074/jbc.M111.254987. View

14.

Suetake I, Shinozaki F, Miyagawa J, Takeshima H, Tajima S . DNMT3L stimulates the DNA methylation activity of Dnmt3a and Dnmt3b through a direct interaction. J Biol Chem. 2004; 279(26):27816-23. DOI: 10.1074/jbc.M400181200. View

15.

Gowher H, Loutchanwoot P, Vorobjeva O, Handa V, Jurkowska R, Jurkowski T . Mutational analysis of the catalytic domain of the murine Dnmt3a DNA-(cytosine C5)-methyltransferase. J Mol Biol. 2006; 357(3):928-41. DOI: 10.1016/j.jmb.2006.01.035. View

16.

Hata K, Okano M, Lei H, Li E . Dnmt3L cooperates with the Dnmt3 family of de novo DNA methyltransferases to establish maternal imprints in mice. Development. 2002; 129(8):1983-93. DOI: 10.1242/dev.129.8.1983. View

17.

Ley T, Ding L, Walter M, McLellan M, Lamprecht T, Larson D . DNMT3A mutations in acute myeloid leukemia. N Engl J Med. 2010; 363(25):2424-33. PMC: 3201818. DOI: 10.1056/NEJMoa1005143. View

18.

Jia D, Jurkowska R, Zhang X, Jeltsch A, Cheng X . Structure of Dnmt3a bound to Dnmt3L suggests a model for de novo DNA methylation. Nature. 2007; 449(7159):248-51. PMC: 2712830. DOI: 10.1038/nature06146. View

19.

Holz-Schietinger C, Matje D, Reich N . Mutations in DNA methyltransferase (DNMT3A) observed in acute myeloid leukemia patients disrupt processive methylation. J Biol Chem. 2012; 287(37):30941-51. PMC: 3438927. DOI: 10.1074/jbc.M112.366625. View

20.

Holz-Schietinger C, Matje D, Harrison M, Reich N . Oligomerization of DNMT3A controls the mechanism of de novo DNA methylation. J Biol Chem. 2011; 286(48):41479-41488. PMC: 3308859. DOI: 10.1074/jbc.M111.284687. View