Abstract
The MLL gene is a common target of chromosomal translocations found in human leukemia. MLL-fusion leukemia has a consistently poor outcome. One of the most common translocation partners is AF9 (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 nuclear magnetic resonance (NMR) solution structure of a DOT1L-AF9 complex. We generate structure-guided point mutations and find that they have graded effects on recruitment of DOT1L to MLL-AF9. Chromatin immunoprecipitation sequencing (ChIP-seq) analyses of H3K79me2 and H3K79me3 show that graded reduction of the DOT1L interaction with MLL-AF9 results in differential loss of H3K79me2 and me3 at MLL-AF9 target genes. Furthermore, the degree of DOT1L recruitment is linked to the level of MLL-AF9 hematopoietic transformation.
| Original language | English (US) |
|---|---|
| Pages (from-to) | 808-820 |
| Number of pages | 13 |
| Journal | Cell reports |
| Volume | 11 |
| Issue number | 5 |
| DOIs | |
| State | Published - 2015 |
Funding
This work was supported by an American Heart Association Predoctoral Fellowship to A.K., an NIH T32 GM080186 Predoctoral Fellowship to A.K., and a grant from the National Cancer Institute (R01 CA155328) to J.H.B., C.S.H., and N.J.Z.-L. This work was also supported by NMR equipment purchased with an NIH High End Instrumentation grant (S10 RR023035) and housed in the Biomolecular Magnetic Resonance Facility at the University of Virginia. We acknowledge expert technical assistance of Shubin Zhang and thank Stefan Bekiranov for helpful advice and discussions.
ASJC Scopus subject areas
- General Biochemistry, Genetics and Molecular Biology
