Journal
JOURNAL OF BIOLOGICAL CHEMISTRY
Volume 297, Issue 5, Pages -Publisher
ELSEVIER
DOI: 10.1016/j.jbc.2021.101288
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Funding
- Agence Nationale de la Recherche (ANR) [ANR-19-CE11-0003-02, ANR-PRCI-19-CE12-0029-01, ANR-20-CE12-0017-03, NIH R01 GM131626]
- NSF [1933344]
- NIH [R01 GM110064, GM136974]
- CNRS
- INSERM
- Strasbourg University
- Investissements d'Avenir grants [ANR-10-IDEX-0002-02, ANR-10-LABX-0030-INRT]
- Directorate For Engineering
- Emerging Frontiers & Multidisciplinary Activities [1933344] Funding Source: National Science Foundation
- Agence Nationale de la Recherche (ANR) [ANR-19-CE11-0003, ANR-20-CE12-0017] Funding Source: Agence Nationale de la Recherche (ANR)
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TFIID, a human general transcription factor, consists of TBP and 13 TAFs, playing a crucial role in Pol II transcription. Assembly of a 5TAF core complex in vitro serves as a building block for forming lobe A or B in TFIID. TAF8 contains distinct regions critical for assembling with the 5TAF core complex in lobe B and interacting with TAF2 in lobe C, both essential for mouse ESC survival.
The human general transcription factor TFIID is composed of the TATA-binding protein (TBP) and 13 TBP-associated factors (TAFs). In eukaryotic cells, TFIID is thought to nucleate RNA polymerase II (Pol II) preinitiation complex formation on all protein coding gene promoters and thus, be crucial for Pol II transcription. TFIID is composed of three lobes, named A, B, and C. A 5TAF core complex can be assembled in vitro constituting a building block for the further assembly of either lobe A or B in TFIID. Structural studies showed that TAF8 forms a histone fold pair with TAF10 in lobe B and participates in connecting lobe B to lobe C. To better understand the role of TAF8 in TFIID, we have investigated the requirement of the different regions of TAF8 for the in vitro assembly of lobe B and C and the importance of certain TAF8 regions for mouse embryonic stem cell (ESC) viability. We have identified a region of TAF8 distinct from the histone fold domain important for assembling with the 5TAF core complex in lobe B. We also delineated four more regions of TAF8 each individually required for interacting with TAF2 in lobe C. Moreover, CRISPR/Cas9-mediated gene editing indicated that the 5TAF core-interacting TAF8 domain and the proline-rich domain of TAF8 that interacts with TAF2 are both required for mouse embryonic stem cell survival. Thus, our study defines distinct TAF8 regions involved in connecting TFIID lobe B to lobe C that appear crucial for TFIID function and consequent ESC survival.
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