c-Myb acetylation at the carboxyl-terminal conserved domain by transcriptional co-activator p300.
Transcription factor c-Myb plays important roles in cell survival and differentiation in immature hematopoietic cells. Here we demonstrate that c-Myb is acetylated at the carboxyl-terminal conserved domain by histone acetyltransferase p300 both in vitro and in vivo. The acetylation sites in vivo have been located at the lysine residues of the conserved domain (K471, K480, K485) by the use of the mutant Myb (Myb-KAmut), in which all three lysine residues are substituted into alanine. Electrophoretic mobility shift assay reveals that Myb-KAmut shows higher DNA binding activity than wild type c-Myb and that acetylation of c-Myb in vitro by p300 causes dramatic increase in DNA binding activity. Accordingly, transactivation activity of both mim-1 and CD34 promoters by Myb-KAmut is higher than that driven by wild type c-Myb. Furthermore, the bromodomain of p300, in addition to the histone acetyltransferase (HAT) domain, is required for effective acetylation of c-Myb, and hGCN5 is revealed to be a factor acetyl-transferase for c-Myb in vitro. We present a new manner of post-translational modification of the c-Myb protein and the potential significance of the acetylation in c-Myb.
Tomita
A
A
First Department of Internal Medicine, Nagoya University School of Medicine, Japan.
Towatari
M
M
Tsuzuki
S
S
Hayakawa
F
F
Kosugi
H
H
Tamai
K
K
Miyazaki
T
T
Kinoshita
T
T
Saito
H
H
D000107:Acetylation; D000123:Acetyltransferases / Q000502:physiology*; D000595:Amino Acid Sequence; D000818:Animals; D018797:Cell Cycle Proteins / Q000502:physiology*; D002460:Cell Line; D017124:Conserved Sequence; D004247:DNA / Q000378:metabolism; D051548:Histone Acetyltransferases; D006801:Humans; D008969:Molecular Sequence Data; D020598:Proto-Oncogene Proteins c-myb / Q000378:metabolism*; D011817:Rabbits; D029701:Saccharomyces cerevisiae Proteins; D015534:Trans-Activators / Q000502:physiology*; D014157:Transcription Factors; D050880:p300-CBP Transcription Factors; D000099224:p300-CBP-Associated Factor
D000123:Acetyltransferases; D018797:Cell Cycle Proteins; D004247:DNA; D051548:Histone Acetyltransferases; D020598:Proto-Oncogene Proteins c-myb; D029701:Saccharomyces cerevisiae Proteins; D015534:Trans-Activators; D014157:Transcription Factors; D050880:p300-CBP Transcription Factors; D000099224:p300-CBP-Associated Factor