The Pol II preinitiation complex (PIC) influences Mediator binding but not promoter-enhancer looping.
Mediator
TFIID
chromatin looping
enhancer
gene expression
promoter
Journal
Genes & development
ISSN: 1549-5477
Titre abrégé: Genes Dev
Pays: United States
ID NLM: 8711660
Informations de publication
Date de publication:
01 08 2021
01 08 2021
Historique:
received:
18
03
2021
accepted:
24
06
2021
pubmed:
25
7
2021
medline:
5
4
2022
entrez:
24
7
2021
Statut:
ppublish
Résumé
Knowledge of how Mediator and TFIID cross-talk contributes to promoter-enhancer (P-E) communication is important for elucidating the mechanism of enhancer function. We conducted an shRNA knockdown screen in murine embryonic stem cells to identify the functional overlap between Mediator and TFIID subunits on gene expression. Auxin-inducible degrons were constructed for TAF12 and MED4, the subunits eliciting the greatest overlap. Degradation of TAF12 led to a dramatic genome-wide decrease in gene expression accompanied by destruction of TFIID, loss of Pol II preinitiation complex (PIC) at promoters, and significantly decreased Mediator binding to promoters and enhancers. Interestingly, loss of the PIC elicited only a mild effect on P-E looping by promoter capture Hi-C (PCHi-C). Degradation of MED4 had a minor effect on Mediator integrity but led to a consistent twofold loss in gene expression, decreased binding of Pol II to Mediator, and decreased recruitment of Pol II to the promoters, but had no effect on the other PIC components. PCHi-C revealed no consistent effect of MED4 degradation on P-E looping. Collectively, our data show that TAF12 and MED4 contribute mechanistically in different ways to P-E communication but neither factor appears to directly control P-E looping, thereby dissociating P-E communication from physical looping.
Identifiants
pubmed: 34301767
pii: gad.348471.121
doi: 10.1101/gad.348471.121
pmc: PMC8336890
doi:
Substances chimiques
Mediator Complex
0
Transcription Factor TFIID
0
RNA Polymerase II
EC 2.7.7.-
Types de publication
Journal Article
Research Support, N.I.H., Extramural
Research Support, Non-U.S. Gov't
Langues
eng
Sous-ensembles de citation
IM
Pagination
1175-1189Subventions
Organisme : NCI NIH HHS
ID : P30 CA016042
Pays : United States
Organisme : NIGMS NIH HHS
ID : R01 GM074701
Pays : United States
Organisme : NIGMS NIH HHS
ID : T32 GM007185
Pays : United States
Informations de copyright
© 2021 Sun et al.; Published by Cold Spring Harbor Laboratory Press.
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