Abstract
AML1 (RUNX1) is one of the most frequently disrupted genes in human leukemias. AML1 encodes transcription factors, which play a pivotal role in hematopoietic differentiation, and their inappropriate expression is associated with leukemic transformation of hematopoietic cells. Previous studies demonstrated that the transcription cofactor p300 binds to the C-terminal region of AML1 and stimulates AML1-dependent transcription during myeloid cell differentiation. Here, we report that AML1 is specifically acetylated by p300 in vitro. Mutagenesis analyses reveal that p300 acetylates AML1 at the two conserved lysine residues (Lys-24 and Lys-43). AML1 is subject to acetylation at the same sites in vivo, and p300-mediated acetylation significantly augments the DNA binding activity of AML1. Disruption of these two lysines severely impairs DNA binding of AML1 and reduced the transcriptional activity and the transforming potential of AML1. Taken together, these data indicate that acetylation of AML1 through p300 is a critical manner of posttranslational modification and identify a novel mechanism for regulating the function of AML1.
Publication types
-
Research Support, Non-U.S. Gov't
MeSH terms
-
Acetylation
-
Animals
-
COS Cells
-
Cell Differentiation
-
Cell Line
-
Cell Line, Tumor
-
Core Binding Factor Alpha 2 Subunit
-
DNA / chemistry
-
DNA Mutational Analysis
-
DNA-Binding Proteins / biosynthesis*
-
DNA-Binding Proteins / genetics*
-
E1A-Associated p300 Protein
-
Gene Expression Regulation*
-
Glutathione Transferase / metabolism
-
HeLa Cells
-
Hematopoietic Stem Cells / cytology
-
Humans
-
Immunoblotting
-
Luciferases / metabolism
-
Lysine / chemistry*
-
Mice
-
Models, Biological
-
Mutation
-
NIH 3T3 Cells
-
Nuclear Proteins / metabolism*
-
Plasmids / metabolism
-
Precipitin Tests
-
Protein Binding
-
Protein Structure, Tertiary
-
Proto-Oncogene Proteins / biosynthesis*
-
Proto-Oncogene Proteins / genetics*
-
Recombinant Fusion Proteins / metabolism
-
Recombinant Proteins / metabolism
-
Trans-Activators / metabolism*
-
Transcription Factors / biosynthesis*
-
Transcription Factors / genetics*
Substances
-
Core Binding Factor Alpha 2 Subunit
-
DNA-Binding Proteins
-
Nuclear Proteins
-
Proto-Oncogene Proteins
-
RUNX1 protein, human
-
Recombinant Fusion Proteins
-
Recombinant Proteins
-
Runx1 protein, mouse
-
Trans-Activators
-
Transcription Factors
-
DNA
-
Luciferases
-
E1A-Associated p300 Protein
-
Ep300 protein, mouse
-
Glutathione Transferase
-
Lysine