TAF8 regions important for TFIID lobe B assembly or for TAF2 interactions are required for embryonic stem cell survival.
CRISPR/Cas9
TAF8
TATA-binding protein (TBP)
TBP-associated factors (TAFs)
TFIID
embryonic stem cells (ESCs)
function
knock out
structure
viability
Journal
The Journal of biological chemistry
ISSN: 1083-351X
Titre abrégé: J Biol Chem
Pays: United States
ID NLM: 2985121R
Informations de publication
Date de publication:
11 2021
11 2021
Historique:
received:
05
07
2021
revised:
01
10
2021
accepted:
06
10
2021
pubmed:
12
10
2021
medline:
15
12
2021
entrez:
11
10
2021
Statut:
ppublish
Résumé
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.
Identifiants
pubmed: 34634302
pii: S0021-9258(21)01091-7
doi: 10.1016/j.jbc.2021.101288
pmc: PMC8564675
pii:
doi:
Substances chimiques
TAF2 protein, human
0
TAF8 protein, human
0
TATA-Binding Protein Associated Factors
0
Tbn protein, mouse
0
Transcription Factor TFIID
0
Transcription Factors
0
Types de publication
Journal Article
Research Support, N.I.H., Extramural
Research Support, Non-U.S. Gov't
Research Support, U.S. Gov't, Non-P.H.S.
Langues
eng
Sous-ensembles de citation
IM
Pagination
101288Subventions
Organisme : Biotechnology and Biological Sciences Research Council
ID : BB/L01386X/1
Pays : United Kingdom
Organisme : NIGMS NIH HHS
ID : R01 GM110064
Pays : United States
Organisme : NIGMS NIH HHS
ID : R01 GM131626
Pays : United States
Organisme : NIGMS NIH HHS
ID : R01 GM136974
Pays : United States
Informations de copyright
Copyright © 2021 The Authors. Published by Elsevier Inc. All rights reserved.
Déclaration de conflit d'intérêts
Conflict of interest The authors declare that they have no conflicts of interest with the contents of this article.