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Series GSE64365 Query DataSets for GSE64365
Status Public on Apr 30, 2015
Title Degree of Recruitment of DOT1L to MLL-AF9 Defines Level of H3K79 Di- and Tri-methylation on Target Genes and Transformation Potential
Organism Mus musculus
Experiment type Genome binding/occupancy profiling by high throughput sequencing
Summary The MLL gene is a common target of chromosomal translocations found in human leukemia. MLL-fusion leukemias are consistently poor prognosis. One of the most common translocation partners is AF9 (a.k.a. MLLT3). MLL-AF9 recruits DOT1L, a histone 3 lysine 79 methyltransferase (H3K79me1/me2/me3), leading to aberrant gene transcription. We show that DOT1L has three AF9 binding sites, and present the NMR solution structure of a DOT1L-AF9 complex. We generated structure-guided point mutations with graded effects on recruitment of DOT1L to MLL-AF9. ChIP-Seq analyses of H3K79me2 and H3K79me3 show that graded reduction of the DOT1L interaction with MLL-AF9 results in selective losses in H3K79me2 and me3 marks at MLL-AF9 target genes. Furthermore, the degree of DOT1L recruitment defines the level of MLL-AF9 hematopoietic transformation.
Overall design Hematopoietic progenitor cells isolated from mouse bone marrow were transduced with retrovirus expressing either wildtype MLL-AF9 (WT), mutants, MLL-AF9 (D544R) and MLL-AF9 (D546R). ChIP-Seq analyses were performed on these wildtype and mutant cells using H3K79me2 and H3K79me3 antibodies. 3 samples corresponding to ChIP-Seq with H3K79me2 antibody: 1) MLL-AF9 (WT) 2) MLL-AF9 (D544R) 3) MLL-AF9 (D546R) 3 Samples Corresponding to ChIP-Seq with H3K79me3 antibody: 4) MLL-AF9 (WT) 5) MLL-AF9 (D544R) 6) MLL-AF9 (D546R)
Contributor(s) Kuntimaddi A, Thorpe J, Singh R, Adli M, Bushweller J
Citation(s) 25921540
Submission date Dec 19, 2014
Last update date May 15, 2019
Contact name John H Bushweller
Organization name University of Virginia
Department Molecular Physiology and Biological Physics
Street address 1340 Jefferson Park Avenue
City Charlottesville
State/province VA
ZIP/Postal code 22908
Country USA
Platforms (1)
GPL16417 Illumina MiSeq (Mus musculus)
Samples (6)
GSM1569542 WT_H3K79me2
GSM1569543 D544R_H3K79me2
GSM1569544 D546R_H3K79me2
BioProject PRJNA270837
SRA SRP051375

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Supplementary file Size Download File type/resource
GSE64365_WholeGenomeAnalysisReadCounts.txt.gz 640.4 Kb (ftp)(http) TXT
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Raw data are available in SRA
Processed data are available on Series record

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