Volume 2, Issue 6, Pages 863-867 (December 1998) The ATM-Related Cofactor Tra1 Is a Component of the Purified SAGA Complex  Patrick A Grant, David Schieltz, Marilyn G Pray-Grant, John R Yates, Jerry L Workman  Molecular Cell  Volume 2, Issue 6, Pages 863-867 (December 1998) DOI: 10.1016/S1097-2765(00)80300-7

Figure 1 Identification of the p400 Subunit of SAGA Silver stain gel of the SAGA complex. Highly purified SAGA complex from the final Mini Q column (Grant et al. 1998b) was analyzed by silver staining of proteins separated on a 4%–20% SDS polyacrylamide gel. The novel p400 subunit of SAGA and the Spt, Ada, and TAFII subunits are indicated. Molecular Cell 1998 2, 863-867DOI: (10.1016/S1097-2765(00)80300-7)

Figure 2 Tra1 Is the p400 Component of SAGA Sequence of the yeast Tra1 protein. The p400 subunit of SAGA was identified by mass spectrometry from 24 peptides, shown in bold. One consecutive peptide sequence (amino acids 2416–2425) and one overlapping peptide sequence (amino acids 2798–2807) are underlined. The region of Tra1 with homology to PI 3-kinases is boxed. Molecular Cell 1998 2, 863-867DOI: (10.1016/S1097-2765(00)80300-7)

Figure 3 Tra1 Stably Associates with SAGA (A) Fluorogram of HAT assays and immunoblots performed with purified SAGA. Fractions eluted from the final Mini Q column were incubated with nucleosomes and assayed for HAT activity or by Western blotting with the indicated antisera. The position of histones H3 and H4, as determined by Coomassie staining of the same gel, are indicated. (B) Immunoprecipitation of the SAGA complex. Partially purified SAGA was incubated with preimmune, anti-Spt8, Tra1, Ada2, and TAFII25 antisera immobilized on protein A–Sepharose beads. A fluorogram of HAT reactions performed with free histones and input material or beads of each antiserum is shown. Molecular Cell 1998 2, 863-867DOI: (10.1016/S1097-2765(00)80300-7)

Figure 4 Tra1 Coelutes with Multiple Protein Complexes HAT assays and immunoblotting performed with fractions from a MonoQ column run with nickel-NTA-agarose-bound sample. The brackets above the lanes indicate the elution of the individual Ada, NuA4, NuA3, and SAGA HAT complexes. Western blotting was performed with anti-Tra1 and Spt8 antisera. Molecular Cell 1998 2, 863-867DOI: (10.1016/S1097-2765(00)80300-7)