TAF8 regions important for TFIID lobe B assembly or for TAF2 interactions are required for embryonic stem cell survival

Elisabeth Scheer, Jie Luo, Andrea Bernardini, Frank Ruffenach, Jean-Marie Garnier, Isabelle Kolb-Cheynel, Kapil Gupta, Imre Berger, Jeff Ranish, László Tora*

*Corresponding author for this work

Research output: Contribution to journalArticle (Academic Journal)peer-review

2 Citations (Scopus)
60 Downloads (Pure)

Abstract

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.

Original languageEnglish
Article number101288
Number of pages15
JournalJournal of Biological Chemistry
Volume297
Issue number5
Early online date9 Oct 2021
DOIs
Publication statusPublished - 1 Nov 2021

Bibliographical note

Funding Information:
We are grateful to the IGBMC cell culture and flow cytometry services for assistance. We thank D. Schmit for starting the initial experiments on this project, F. El-Saafin and the Tora lab members for helpful discussions, N. Jung and B. Reina San Martin for help with the CRISPR/Cas9 experiments, and L. Negroni with MS experiments and data submission. This study was supported by grants from 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 and NSF (Award Number: 1933344) grants (to L. T.); NIH R01 GM110064 and GM136974 (to J. R.) and supported by funds from CNRS, INSERM, Strasbourg University, and Investissements d?Avenir grants (ANR-10-IDEX-0002-02 and ANR-10-LABX-0030-INRT). The content is solely the responsibility of the authors and does not necessarily represent the official views of the National Institutes of Health or other granting agencies.

Funding Information:
Funding and additional information—This study was supported by grants from 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 and NSF (Award Number: 1933344) grants (to L. T.); NIH R01 GM110064 and GM136974 (to J. R.) and supported by funds from CNRS, INSERM, Strasbourg University, and Investissements d’Avenir grants (ANR-10-IDEX-0002-02 and ANR-10-LABX-0030-INRT). The content is solely the responsibility of the authors and does not necessarily represent the official views of the National Institutes of Health or other granting agencies.

Publisher Copyright:
© 2021 THE AUTHORS. Published by Elsevier Inc on behalf of American Society for Biochemistry and Molecular Biology.

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