@article{6b745e95e90248b195bb1ebbd0e566c6,
title = "DOT1L activity in leukemia cells requires interaction with ubiquitylated H2B that promotes productive nucleosome binding",
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.",
author = "Spangler, {Cathy J.} and Yadav, {Satya P.} and Dongxu Li and Geil, {Carinne N.} and Smith, {Charlotte B.} and Wang, {Gang Greg} and Lee, {Tae Hee} and McGinty, {Robert K.}",
note = "Funding Information: MV4; 11 cells bearing inducible Cas9 (MV4; 11-iCas9) were a kind gift of Drs. X. Shi and H. Wen. We thank UNC's facilities, including the Flow Cytometry Core and the Tissue Culture Facility, for their professional assistance with this work. We also thank the McGinty, Lee, and Wang laboratories for helpful discussion and critical comments on the manuscript. This work was funded by NIH grants ( R35GM133498 to R.K.M.; F99CA253730 to C.J.S.; R01GM123164 and R01GM130793 to T.-H.L.; R01CA211336 and R01CA215284 to G.G.W.). The cores affiliated to UNC Cancer Center are supported in part by the UNC Lineberger Comprehensive Cancer Center Core Support Grant P30CA016086 . G.G.W. is an American Cancer Society (ACS) Research Scholar and a Leukemia and Lymphoma Society (LLS) Scholar. R.K.M. is a Searle Scholar and a Pew-Stewart Scholar for Cancer Research. Graphical abstract was created in part with BioRender . Funding Information: MV4; 11 cells bearing inducible Cas9 (MV4; 11-iCas9) were a kind gift of Drs. X. Shi and H. Wen. We thank UNC's facilities, including the Flow Cytometry Core and the Tissue Culture Facility, for their professional assistance with this work. We also thank the McGinty, Lee, and Wang laboratories for helpful discussion and critical comments on the manuscript. This work was funded by NIH grants (R35GM133498 to R.K.M.; F99CA253730 to C.J.S.; R01GM123164 and R01GM130793 to T.-H.L.; R01CA211336 and R01CA215284 to G.G.W.). The cores affiliated to UNC Cancer Center are supported in part by the UNC Lineberger Comprehensive Cancer Center Core Support Grant P30CA016086. G.G.W. is an American Cancer Society (ACS) Research Scholar and a Leukemia and Lymphoma Society (LLS) Scholar. R.K.M. is a Searle Scholar and a Pew-Stewart Scholar for Cancer Research. Graphical abstract was created in part with BioRender. C.J.S. prepared proteins and nucleosomes for biochemical studies, performed bulk fluorescence experiments, and prepared constructs for cellular studies. S.P.Y. prepared nucleosomes and performed single-molecule measurements. D.L. performed cellular studies. C.N.G. and C.B.S. prepared proteins for biochemical studies. G.G.W. T.-H.L. and R.K.M. provided funding and supervised the studies. C.J.S. D.L. S.P.Y. G.G.W. T.-H.L. and R.K.M. conceived the studies, analyzed the data, and prepared the manuscript with contributions from all authors. The authors declare no competing interests. Publisher Copyright: {\textcopyright} 2022 The Authors",
year = "2022",
month = feb,
day = "15",
doi = "10.1016/j.celrep.2022.110369",
language = "English (US)",
volume = "38",
journal = "Cell Reports",
issn = "2211-1247",
publisher = "Cell Press",
number = "7",
}