Abstract
The roles of RNA 5-methylcytosine (RNA:m5C) and RNA:m5C methyltransferases (RCMTs) in lineage-associated chromatin organization and drug response/resistance are unclear. Here we demonstrate that the RCMTs, namely NSUN3 and DNMT2, directly bind hnRNPK, a conserved RNA-binding protein. hnRNPK interacts with the lineage-determining transcription factors (TFs), GATA1 and SPI1/PU.1, and with CDK9/P-TEFb to recruit RNA-polymerase-II at nascent RNA, leading to formation of 5-Azacitidine (5-AZA)-sensitive chromatin structure. In contrast, NSUN1 binds BRD4 and RNA-polymerase-II to form an active chromatin structure that is insensitive to 5-AZA, but hypersensitive to the BRD4 inhibitor JQ1 and to the downregulation of NSUN1 by siRNAs. Both 5-AZA-resistant leukaemia cell lines and clinically 5-AZA-resistant myelodysplastic syndrome and acute myeloid leukaemia specimens have a significant increase in RNA:m5C and NSUN1-/BRD4-associated active chromatin. This study reveals novel RNA:m5C/RCMT-mediated chromatin structures that modulate 5-AZA response/resistance in leukaemia cells, and hence provides a new insight into treatment of leukaemia.
Publication types
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Research Support, Non-U.S. Gov't
MeSH terms
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Antineoplastic Agents / pharmacology*
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Azacitidine / pharmacology*
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Bone Marrow / drug effects
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Bone Marrow / metabolism
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Bone Marrow / pathology
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Cell Cycle Proteins
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Cell Line, Tumor
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Chromatin / chemistry
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Chromatin / drug effects
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Chromatin / metabolism
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Chromatin Assembly and Disassembly
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Cyclin-Dependent Kinase 9 / genetics
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Cyclin-Dependent Kinase 9 / metabolism
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Cytosine / metabolism
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DNA (Cytosine-5-)-Methyltransferases / genetics
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DNA (Cytosine-5-)-Methyltransferases / metabolism
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Drug Resistance, Neoplasm / genetics*
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GATA1 Transcription Factor / genetics
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GATA1 Transcription Factor / metabolism
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Gene Expression Regulation, Leukemic*
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Heterogeneous-Nuclear Ribonucleoprotein K / genetics
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Heterogeneous-Nuclear Ribonucleoprotein K / metabolism
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Humans
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Leukemia, Myeloid, Acute / drug therapy
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Leukemia, Myeloid, Acute / genetics*
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Leukemia, Myeloid, Acute / metabolism
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Leukemia, Myeloid, Acute / pathology
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Methylation
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Methyltransferases / genetics
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Methyltransferases / metabolism
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Myelodysplastic Syndromes / drug therapy
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Myelodysplastic Syndromes / genetics*
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Myelodysplastic Syndromes / metabolism
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Myelodysplastic Syndromes / pathology
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Nuclear Proteins / genetics
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Nuclear Proteins / metabolism
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Protein Binding
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Proto-Oncogene Proteins / genetics
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Proto-Oncogene Proteins / metabolism
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RNA, Neoplasm / genetics*
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RNA, Neoplasm / metabolism
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RNA, Small Interfering / genetics
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RNA, Small Interfering / metabolism
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Signal Transduction
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Trans-Activators / genetics
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Trans-Activators / metabolism
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Transcription Factors / genetics
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Transcription Factors / metabolism
Substances
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Antineoplastic Agents
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BRD4 protein, human
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Cell Cycle Proteins
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Chromatin
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GATA1 Transcription Factor
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GATA1 protein, human
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Heterogeneous-Nuclear Ribonucleoprotein K
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Nuclear Proteins
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Proto-Oncogene Proteins
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RNA, Neoplasm
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RNA, Small Interfering
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Trans-Activators
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Transcription Factors
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proto-oncogene protein Spi-1
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HNRNPK protein, human
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Cytosine
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Methyltransferases
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NSUN2 protein, human
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NSUN3 protein, human
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DNA (Cytosine-5-)-Methyltransferases
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TRDMT1 protein, human
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CDK9 protein, human
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Cyclin-Dependent Kinase 9
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Azacitidine