DOT1L activity in leukemia cells requires interaction with ubiquitylated H2B that promotes productive nucleosome binding

Cathy J Spangler; Satya P Yadav; Dongxu Li; Carinne N Geil; Charlotte B Smith; Gang Greg Wang; Tae-Hee Lee; Robert K McGinty

doi:10.1016/j.celrep.2022.110369

DOT1L activity in leukemia cells requires interaction with ubiquitylated H2B that promotes productive nucleosome binding

Cell Rep. 2022 Feb 15;38(7):110369. doi: 10.1016/j.celrep.2022.110369.

Authors

Cathy J Spangler¹, Satya P Yadav², Dongxu Li³, Carinne N Geil⁴, Charlotte B Smith⁴, Gang Greg Wang⁵, Tae-Hee Lee⁶, Robert K McGinty⁷

Affiliations

¹ Department of Biochemistry and Biophysics, School of Medicine, University of North Carolina at Chapel Hill, Chapel Hill, NC 27599, USA.
² Department of Chemistry, Pennsylvania State University, University Park, PA 16802, USA.
³ Department of Biochemistry and Biophysics, School of Medicine, University of North Carolina at Chapel Hill, Chapel Hill, NC 27599, USA; Lineberger Comprehensive Cancer Center, University of North Carolina at Chapel Hill, Chapel Hill, NC 27599, USA.
⁴ Division of Chemical Biology and Medicinal Chemistry, Eshelman School of Pharmacy, University of North Carolina at Chapel Hill, Chapel Hill, NC 27599, USA.
⁵ Department of Biochemistry and Biophysics, School of Medicine, University of North Carolina at Chapel Hill, Chapel Hill, NC 27599, USA; Lineberger Comprehensive Cancer Center, University of North Carolina at Chapel Hill, Chapel Hill, NC 27599, USA; Department of Pharmacology, School of Medicine, University of North Carolina at Chapel Hill, Chapel Hill, NC 27599, USA. Electronic address: [email protected].
⁶ Department of Chemistry, Pennsylvania State University, University Park, PA 16802, USA; Huck Institutes of the Life Sciences, Pennsylvania State University, University Park, PA 16802, USA. Electronic address: [email protected].
⁷ Department of Biochemistry and Biophysics, School of Medicine, University of North Carolina at Chapel Hill, Chapel Hill, NC 27599, USA; Lineberger Comprehensive Cancer Center, University of North Carolina at Chapel Hill, Chapel Hill, NC 27599, USA; Division of Chemical Biology and Medicinal Chemistry, Eshelman School of Pharmacy, University of North Carolina at Chapel Hill, Chapel Hill, NC 27599, USA. Electronic address: [email protected].

Abstract

DOT1L methylates histone H3 lysine 79 during transcriptional elongation and is stimulated by ubiquitylation of histone H2B lysine 120 (H2BK120ub) in a classical trans-histone crosstalk pathway. Aberrant genomic localization of DOT1L is implicated in mixed lineage leukemia (MLL)-rearranged leukemias, an aggressive subset of leukemias that lacks effective targeted treatments. Despite recent atomic structures of DOT1L in complex with H2BK120ub nucleosomes, fundamental questions remain as to how DOT1L-ubiquitin and DOT1L-nucleosome acidic patch interactions observed in these structures contribute to nucleosome binding and methylation by DOT1L. Here, we combine bulk and single-molecule biophysical measurements with cancer cell biology to show that ubiquitin and cofactor binding drive conformational changes to stimulate DOT1L activity. Using structure-guided mutations, we demonstrate that ubiquitin and nucleosome acidic patch binding by DOT1L are required for cell proliferation in the MV4; 11 leukemia model, providing proof of principle for MLL targeted therapeutic strategies.

Keywords: DOT1L; MLL-rearranged leukemia; chromatin; histone; lysine methyltransferase; mixed lineage leukemia (MLL); nucleosome; single-molecule FRET.

Publication types

Research Support, N.I.H., Extramural
Research Support, Non-U.S. Gov't

MeSH terms

Cell Line, Tumor
Cell Proliferation
Gene Rearrangement
Histone-Lysine N-Methyltransferase / chemistry
Histone-Lysine N-Methyltransferase / genetics
Histone-Lysine N-Methyltransferase / metabolism*
Histones / metabolism
Humans
Leukemia / metabolism*
Leukemia / pathology
Lysine / metabolism
Male
Methylation
Models, Molecular
Myeloid-Lymphoid Leukemia Protein / genetics
Nucleosomes / metabolism*
Protein Binding
Ubiquitin / metabolism
Ubiquitination*

Substances

Histones
KMT2A protein, human
Nucleosomes
Ubiquitin
Myeloid-Lymphoid Leukemia Protein
DOT1L protein, human
Histone-Lysine N-Methyltransferase
Lysine

Abstract

Publication types

MeSH terms

Substances

Grants and funding