TY  - JOUR
A1  - Klein, Joachim
A1  - Nolden, Mark
A1  - Sanders, Steven L.
A1  - Kirchner, Jay
A1  - Weil, P. Anthony
A1  - Melcher, Karsten
T1  - Use of a genetically introduced cross-linker to identify interaction sites of acidic activators within native transcription factor IID and SAGA
T2  - Journal of biological chemistry
N2  - An important goal is to identify the direct activation domain (AD)-interacting components of the transcriptional machinery within the context of native complexes. Toward this end, we first demonstrate that the multisubunit TFIID, SAGA, mediator, and Swi/Snf coactivator complexes from transcriptionally competent whole-cell yeast extracts were all capable of specifically interacting with the prototypic acidic ADs of Gal4 and VP16. We then used hexahistidine tags as genetically introduced activation domain-localized cross-linking receptors. In combination with immunological reagents against all subunits of TFIID and SAGA, we systematically identified the direct AD-interacting subunits within the AD-TFIID and AD-SAGA coactivator complexes enriched from whole-cell extracts and confirmed these results using purified TFIID and partially purified SAGA. Both ADs directly cross-linked to TBP and to a subset of TFIID and SAGA subunits that carry histone-fold motifs.
Y1  - 2021
UR  - http://publikationen.ub.uni-frankfurt.de/frontdoor/index/index/docId/76088
UR  - https://nbn-resolving.org/urn:nbn:de:hebis:30:3-760886
SN  - 0021-9258
VL  - 278.2003
IS  - 9
SP  - 6779
EP  - 6786
PB  - American Society for Biochemistry and Molecular Biology Publications
CY  - Bethesda, Md
ER  -