Characterization of the DOT1L network: implications of diverse roles for DOT1L.
Methylation of lysine 79 on histone H3 (H3K79) is mediated by a methyltransferase called Dot1-like protein (DOT1L). DOT1L is involved in the regulation of telomeric silencing, development, cell cycle checkpoint and transcription. However, the mechanisms by which DOT1L controls these unrelated and diverse functions are unknown. To gain greater insight into DOT1L-mediated functions, we have purified a DOT1L-containing complex using tandem affinity purification. Mass spectrometry of the DOT1L-containing complex revealed that AF9, ENL and NPM1 were shown to be major DOT1L-interacting proteins. To construct a plausible DOT1L-interaction web, AF9-, ENL- and NPM1-containing complexes were also purified for mass spectrometry analysis. The data showed that DOT1L might control AF9- and ENL-mediated transcription, regulate RNA processing, and function as a histone chaperone in a NPM1-dependent manner. In addition, the purification of protein complexes identified a number of novel interacting partners associated with DOT1L, AF9, ENL and NPM1. These data define a unique DOT1L network and shed light on unknown functions of the DOT1L complex.
Park
Geunyeong
G
Department of Pharmacology, School of Medicine, Kyung Hee University, 1 Hoegi-Dong, Dongdaemun-Gu, Seoul 130-701, Republic of Korea.
Gong
Zihua
Z
Chen
Junjie
J
Kim
Ja-Eun
JE
D000818:Animals; D015153:Blotting, Western; D002460:Cell Line; D004268:DNA-Binding Proteins / Q000737:chemistry / Q000235:genetics / Q000378:metabolism*; D006657:Histones / Q000378:metabolism; D006801:Humans; D008780:Methyltransferases / Q000737:chemistry / Q000235:genetics / Q000378:metabolism*; D009687:Nuclear Proteins / Q000737:chemistry / Q000235:genetics / Q000378:metabolism*; D000090243:Nucleophosmin; D025941:Protein Interaction Mapping / Q000379:methods*; D011994:Recombinant Proteins / Q000737:chemistry / Q000235:genetics / Q000378:metabolism; D014157:Transcription Factors / Q000737:chemistry / Q000235:genetics / Q000378:metabolism*; D014158:Transcription, Genetic
D004268:DNA-Binding Proteins; D006657:Histones; C000716967:NPM1 protein, human; D009687:Nuclear Proteins; D011994:Recombinant Proteins; D014157:Transcription Factors; D000090243:Nucleophosmin; D008780:Methyltransferases