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Structure paper

TitleArchitecture of the yeast Pol III pre-termination complex and pausing mechanism on poly(dT) termination signals.
Journal, issue, pagesCell Rep, Vol. 40, Issue 10, Page 111316, Year 2022
Publish dateSep 6, 2022
AuthorsMathias Girbig / Juanjuan Xie / Helga Grötsch / Domenico Libri / Odil Porrua / Christoph W Müller /
PubMed AbstractRNA polymerase (Pol) III is specialized to transcribe short, abundant RNAs, for which it terminates transcription on polythymine (dT) stretches on the non-template (NT) strand. When Pol III reaches ...RNA polymerase (Pol) III is specialized to transcribe short, abundant RNAs, for which it terminates transcription on polythymine (dT) stretches on the non-template (NT) strand. When Pol III reaches the termination signal, it pauses and forms the pre-termination complex (PTC). Here, we report cryoelectron microscopy (cryo-EM) structures of the yeast Pol III PTC and complementary functional states at resolutions of 2.7-3.9 Å. Pol III recognizes the poly(dT) termination signal with subunit C128 that forms a hydrogen-bond network with the NT strand and, thereby, induces pausing. Mutating key interacting residues interferes with transcription termination in vitro, impairs yeast growth, and causes global termination defects in vivo, confirming our structural results. Additional cryo-EM analysis reveals that C53-C37, a Pol III subcomplex and key termination factor, participates indirectly in Pol III termination. We propose a mechanistic model of Pol III transcription termination and rationalize why Pol III, unlike Pol I and Pol II, terminates on poly(dT) signals.
External linksCell Rep / PubMed:36070694
MethodsEM (single particle)
Resolution2.7 - 3.9 Å
Structure data

14447

7z1l

EMDB-14447, PDB-7z1l:
Structure of yeast RNA Polymerase III Pre-Termination Complex (PTC)
Method: EM (single particle) / Resolution: 2.8 Å

14448

7z1m

EMDB-14448, PDB-7z1m:
Structure of yeast RNA Polymerase III Elongation Complex (EC)
Method: EM (single particle) / Resolution: 3.4 Å

14449

7z1n

EMDB-14449, PDB-7z1n:
Structure of yeast RNA Polymerase III Delta C53-C37-C11
Method: EM (single particle) / Resolution: 3.9 Å

EMDB-14450: Pol III PTC - Full transcription bubble
Method: EM (single particle) / Resolution: 3.2 Å

14451

7z1o

EMDB-14451, PDB-7z1o:
Structure of yeast RNA Polymerase III PTC + NTPs
Method: EM (single particle) / Resolution: 2.7 Å

Chemicals

ChemComp-ZN:
Unknown entry

ChemComp-MG:
Unknown entry

ChemComp-4QM:
(3R,5S,7R,8R,9S,10S,12S,13R,14S,17R)-10,13-dimethyl-17-[(2R)-pentan-2-yl]-2,3,4,5,6,7,8,9,11,12,14,15,16,17-tetradecahydro-1H-cyclopenta[a]phenanthrene-3,7,12-triol

ChemComp-1N7:
CHAPSO / detergent*YM

Source
  • saccharomyces cerevisiae w303 (yeast)
  • synthetic construct (others)
KeywordsTRANSCRIPTION / RNA synthesis / short RNAs / termination

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