Phosphorylation of AML1/RUNX1 regulates its degradation and nuclear matrix association.
The acute myeloid leukemia 1 (AML1) transcription factors are key regulators of hematopoietic differentiation. Cellular AML1c protein is found in the nucleus and can be separated into two fractions, one soluble in buffers containing salt and nonionic detergent and the other insoluble and tightly bound to the nuclear matrix. We find that the AML1c protein is modified by both phosphorylation and ubiquitination. Our studies show that the majority of the ubiquitinated AML1c is associated with the insoluble nuclear matrix. Treatment of cells with the proteasome inhibitor PS341 (Velcade, Bortezomib) increases the levels of ubiquitinated AML1c. Mutation of the four phosphorylation sites necessary for transcriptional regulation (serine 276, serine 293, serine 303, and threonine 300) mimics the effects of the proteasome inhibitor, increasing the levels of ubiquitinated, matrix-bound AML1c. We find that the soluble and insoluble forms of AML1c are degraded at a similar rate. However, mutation of these four serine/threonine residues statistically increases the half-life of the matrix-associated AML1c. Thus, phosphorylation of AML1c on specific serine/threonine residues controls both transcriptional activity and rate of degradation.
Biggs
Joseph R
JR
Department of Medicine, P.O. Box 250955, Medical University of South Carolina, 6 Jonathan Lucas Street, Charleston, SC 29425, USA.
Zhang
Youhong
Y
Peterson
Luke F
LF
Garcia
Marileila
M
Zhang
Dong-Er
DE
Kraft
Andrew S
AS
D000818:Animals; D001897:Boronic Acids / Q000494:pharmacology; D000069286:Bortezomib; D050676:Core Binding Factor Alpha 2 Subunit; D004268:DNA-Binding Proteins / Q000187:drug effects / Q000235:genetics / Q000378:metabolism*; D047468:Immunoprecipitation; D051379:Mice; D018345:Mice, Knockout; D009154:Mutation; D015530:Nuclear Matrix / Q000378:metabolism*; D010766:Phosphorylation; D011480:Protease Inhibitors / Q000494:pharmacology; D011518:Proto-Oncogene Proteins / Q000187:drug effects / Q000235:genetics / Q000378:metabolism*; D011719:Pyrazines / Q000494:pharmacology; D012694:Serine / Q000235:genetics; D013912:Threonine / Q000235:genetics; D014157:Transcription Factors / Q000187:drug effects / Q000235:genetics / Q000378:metabolism*; D014158:Transcription, Genetic; D025801:Ubiquitin / Q000378:metabolism
D001897:Boronic Acids; D050676:Core Binding Factor Alpha 2 Subunit; D004268:DNA-Binding Proteins; D011480:Protease Inhibitors; D011518:Proto-Oncogene Proteins; D011719:Pyrazines; C493726:Runx1 protein, mouse; D014157:Transcription Factors; D025801:Ubiquitin; D013912:Threonine; D012694:Serine; D000069286:Bortezomib