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Structure paper

TitleStructural basis of a transcription pre-initiation complex on a divergent promoter.
Journal, issue, pagesMol Cell, Vol. 83, Issue 4, Page 574-588.e11, Year 2023
Publish dateFeb 16, 2023
AuthorsJose J Gorbea Colón / Leon Palao / Shin-Fu Chen / Hee Jong Kim / Laura Snyder / Yi-Wei Chang / Kuang-Lei Tsai / Kenji Murakami /
PubMed AbstractMost eukaryotic promoter regions are divergently transcribed. As the RNA polymerase II pre-initiation complex (PIC) is intrinsically asymmetric and responsible for transcription in a single ...Most eukaryotic promoter regions are divergently transcribed. As the RNA polymerase II pre-initiation complex (PIC) is intrinsically asymmetric and responsible for transcription in a single direction, it is unknown how divergent transcription arises. Here, the Saccharomyces cerevisiae Mediator complexed with a PIC (Med-PIC) was assembled on a divergent promoter and analyzed by cryoelectron microscopy. The structure reveals two distinct Med-PICs forming a dimer through the Mediator tail module, induced by a homodimeric activator protein localized near the dimerization interface. The tail dimer is associated with ∼80-bp upstream DNA, such that two flanking core promoter regions are positioned and oriented in a suitable form for PIC assembly in opposite directions. Also, cryoelectron tomography visualized the progress of the PIC assembly on the two core promoter regions, providing direct evidence for the role of the Med-PIC dimer in divergent transcription.
External linksMol Cell / PubMed:36731470 / PubMed Central
MethodsEM (single particle) / EM (subtomogram averaging)
Resolution3.3 - 36.0 Å
Structure data

26542

7ui9

EMDB-26542, PDB-7ui9:
Core Mediator-PICearly (Copy A)
Method: EM (single particle) / Resolution: 3.3 Å

26543

7uic

EMDB-26543, PDB-7uic:
Mediator-PIC Early (Tail A)
Method: EM (single particle) / Resolution: 3.7 Å

26544

7uif

EMDB-26544, PDB-7uif:
Mediator-PIC Early (Core B)
Method: EM (single particle) / Resolution: 4.6 Å

26545

7uig

EMDB-26545, PDB-7uig:
Mediator-PIC Early (Mediator A)
Method: EM (single particle) / Resolution: 4.3 Å

26547

7uik

EMDB-26547, PDB-7uik:
Mediator-PIC Early (Tail A + Upstream DNA & Activator)
Method: EM (single particle) / Resolution: 7.7 Å

26548

7uil

EMDB-26548, PDB-7uil:
Mediator-PIC Early (Tail A/B Dimer)
Method: EM (single particle) / Resolution: 4.3 Å

26551

7uio

EMDB-26551, PDB-7uio:
Mediator-PIC Early (Composite Model)
Method: EM (single particle) / Resolution: 3.3 Å

EMDB-28023: Dimerized Mediator-PIC with early GTFs and TFIIE on a divergent promoter
Method: EM (subtomogram averaging) / Resolution: 26.0 Å

EMDB-28024: Dimerized Mediator PIC with early GTFs on a divergent promoter
Method: EM (subtomogram averaging) / Resolution: 36.0 Å

Chemicals

ChemComp-ZN:
Unknown entry

ChemComp-THR:
THREONINE / Threonine

ChemComp-ALA:
ALANINE / Alanine

ChemComp-ASP:
ASPARTIC ACID / Aspartic acid

Source
  • saccharomyces cerevisiae s288c (yeast)
  • saccharomyces cerevisiae (brewer's yeast)
  • baker's yeast (brewer's yeast)
KeywordsTRANSCRIPTION / Divergent transcription / Mediator / RNA Polymerase II / PIC / Pre-Initiation / Activation / Transcription Factor / Gal4 / Gal4VP16

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