Abstract
RUNX1/AML1 is required for the development of definitive hematopoiesis, and its activity is altered by mutations, deletions, and chromosome translocations in human acute leukemia. RUNX1 function can be regulated by post-translational modifications and protein-protein interactions. We show that RUNX1 is arginine-methylated in vivo by the arginine methyltransferase PRMT1, and that PRMT1 serves as a transcriptional coactivator for RUNX1 function. Using mass spectrometry, and a methyl-arginine-specific antibody, we identified two arginine residues (R206 and R210) within the region of RUNX1 that interact with the corepressor SIN3A and are methylated by PRMT1. PRMT1- dependent methylation of RUNX1 at these arginine residues abrogates its association with SIN3A, whereas shRNA against PRMT1 (or use of a methyltransferase inhibitor) enhances this association. We find arginine-methylated RUNX1 on the promoters of two bona fide RUNX1 target genes, CD41 and PU.1 and show that shRNA against PRMT1 or RUNX1 down-regulates their expression. These arginine methylation sites and the dynamic regulation of corepressor binding are lost in the leukemia-associated RUNX1-ETO fusion protein, which likely contributes to its dominant inhibitory activity.
Publication types
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Research Support, N.I.H., Extramural
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Research Support, Non-U.S. Gov't
MeSH terms
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Antigens, CD34 / analysis
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Arginine / analysis
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Arginine / metabolism
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Cell Line, Tumor
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Core Binding Factor Alpha 2 Subunit / chemistry
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Core Binding Factor Alpha 2 Subunit / genetics
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Core Binding Factor Alpha 2 Subunit / metabolism*
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DNA-Binding Proteins / metabolism
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Gene Expression Regulation*
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Hematopoiesis / genetics
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Humans
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Methylation
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Mutation
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Platelet Membrane Glycoprotein IIb / genetics*
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Protein-Arginine N-Methyltransferases / metabolism*
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Proto-Oncogene Proteins / genetics*
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Proto-Oncogene Proteins / metabolism
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RNA, Small Interfering / genetics
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RNA, Small Interfering / pharmacology
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RUNX1 Translocation Partner 1 Protein
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Repressor Proteins / metabolism*
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Sin3 Histone Deacetylase and Corepressor Complex
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Trans-Activators / genetics*
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Transcription Factors / metabolism
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Transcription, Genetic / drug effects
Substances
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Antigens, CD34
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Core Binding Factor Alpha 2 Subunit
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DNA-Binding Proteins
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Platelet Membrane Glycoprotein IIb
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Proto-Oncogene Proteins
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RNA, Small Interfering
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RUNX1 Translocation Partner 1 Protein
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RUNX1 protein, human
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RUNX1T1 protein, human
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Repressor Proteins
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SIN3A transcription factor
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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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Arginine
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PRMT1 protein, human
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Protein-Arginine N-Methyltransferases
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Sin3 Histone Deacetylase and Corepressor Complex