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TitleRegulation of mammalian transcription by Gdown1 through a novel steric crosstalk revealed by cryo-EM.
Journal, issue, pagesEMBO J, Vol. 31, Issue 17, Page 3575-3587, Year 2012
Publish dateAug 29, 2012
AuthorsYi-Min Wu / Jen-Wei Chang / Chun-Hsiung Wang / Yen-Chen Lin / Pei-lun Wu / Shih-hsin Huang / Chia-Chi Chang / Xiaopeng Hu / Averell Gnatt / Wei-hau Chang /
PubMed AbstractIn mammals, a distinct RNA polymerase II form, RNAPII(G) contains a novel subunit Gdown1 (encoded by POLR2M), which represses gene activation, only to be reversed by the multisubunit Mediator co- ...In mammals, a distinct RNA polymerase II form, RNAPII(G) contains a novel subunit Gdown1 (encoded by POLR2M), which represses gene activation, only to be reversed by the multisubunit Mediator co-activator. Here, we employed single-particle cryo-electron microscopy (cryo-EM) to disclose the architectures of RNAPII(G), RNAPII and RNAPII in complex with the transcription initiation factor TFIIF, all to ~19 Å. Difference analysis mapped Gdown1 mostly to the RNAPII Rpb5 shelf-Rpb1 jaw, supported by antibody labelling experiments. These structural features correlate with the moderate increase in the efficiency of RNA chain elongation by RNAP II(G). In addition, our updated RNAPII-TFIIF map showed that TFIIF tethers multiple regions surrounding the DNA-binding cleft, in agreement with cross-linking and biochemical mapping. Gdown1's binding sites overlap extensively with those of TFIIF, with Gdown1 sterically excluding TFIIF from RNAPII, herein demonstrated by competition assays using size exclusion chromatography. In summary, our work establishes a structural basis for Gdown1 impeding initiation at promoters, by obstruction of TFIIF, accounting for an additional dependent role of Mediator in activated transcription.
External linksEMBO J / PubMed:22850672 / PubMed Central
MethodsEM (single particle)
Resolution19.0 Å
Structure data

EMDB-5440:
Regulation of mammalian transcription by Gdown1 through a novel steric crosstalk revealed by cryo-EM
Method: EM (single particle) / Resolution: 19.0 Å

EMDB-5441:
Regulation of mammalian transcription by Gdown1 through a novel steric crosstalk revealed by cryo-EM
Method: EM (single particle) / Resolution: 19.0 Å

EMDB-5442:
Regulation of mammalian transcription by Gdown1 through a novel steric crosstalk revealed by cryo-EM
Method: EM (single particle) / Resolution: 19.0 Å

EMDB-5443:
Regulation of mammalian transcription by Gdown1 through a novel steric crosstalk revealed by cryo-EM
Method: EM (single particle) / Resolution: 19.0 Å

Source
  • Bos taurus (cattle)

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