Host cell factor and an uncharacterized SANT domain protein are stable components of ATAC, a novel dAda2A/dGcn5-containing histone acetyltransferase complex in Drosophila.
Gcn5 is a conserved histone acetyltransferase (HAT) found in a number of multisubunit complexes from Saccharomyces cerevisiae, mammals, and flies. We previously identified Drosophila melanogaster homologues of the yeast proteins Ada2, Ada3, Spt3, and Tra1 and showed that they associate with dGcn5 to form at least two distinct HAT complexes. There are two different Ada2 homologues in Drosophila named dAda2A and dAda2B. dAda2B functions within the Drosophila version of the SAGA complex (dSAGA). To gain insight into dAda2A function, we sought to identify novel components of the complex containing this protein, ATAC (Ada two A containing) complex. Affinity purification and mass spectrometry revealed that, in addition to dAda3 and dGcn5, host cell factor (dHCF) and a novel SANT domain protein, named Atac1 (ATAC component 1), copurify with this complex. Coimmunoprecipitation experiments confirmed that these proteins associate with dGcn5 and dAda2A, but not with dSAGA-specific components such as dAda2B and dSpt3. Biochemical fractionation revealed that ATAC has an apparent molecular mass of 700 kDa and contains dAda2A, dGcn5, dAda3, dHCF, and Atac1 as stable subunits. Thus, ATAC represents a novel histone acetyltransferase complex that is distinct from previously purified Gcn5/Pcaf-containing complexes from yeast and mammalian cells.
Guelman
Sebastián
S
Stowers Institute for Medical Research, 1000 E. 50th St., Kansas City, MO 64110, USA.
Suganuma
Tamaki
T
Florens
Laurence
L
Swanson
Selene K
SK
Kiesecker
Cheri L
CL
Kusch
Thomas
T
Anderson
Scott
S
Yates
John R
JR
Washburn
Michael P
MP
Abmayr
Susan M
SM
Workman
Jerry L
JL
D000595:Amino Acid Sequence; D000818:Animals; D000906:Antibodies / Q000276:immunology; D002875:Chromosomes / Q000737:chemistry; D029721:Drosophila Proteins / Q000032:analysis / Q000276:immunology / Q000378:metabolism*; D004331:Drosophila melanogaster / Q000378:metabolism*; D051548:Histone Acetyltransferases / Q000032:analysis / Q000378:metabolism*; D047468:Immunoprecipitation; D008969:Molecular Sequence Data; D000099224:p300-CBP-Associated Factor
D000906:Antibodies; D029721:Drosophila Proteins; D051548:Histone Acetyltransferases; C470558:Ada2a protein, Drosophila; C508027:Atac1 protein, Drosophila; C473147:host cell factor, Drosophila; C470559:ADA3 protein, Drosophila; C470557:GCN5 protein, Drosophila; D000099224:p300-CBP-Associated Factor