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Structure paper

TitleIn-cell architecture of an actively transcribing-translating expressome.
Journal, issue, pagesScience, Vol. 369, Issue 6503, Page 554-557, Year 2020
Publish dateJul 31, 2020
AuthorsFrancis J O'Reilly / Liang Xue / Andrea Graziadei / Ludwig Sinn / Swantje Lenz / Dimitry Tegunov / Cedric Blötz / Neil Singh / Wim J H Hagen / Patrick Cramer / Jörg Stülke / Julia Mahamid / Juri Rappsilber /
PubMed AbstractStructural biology studies performed inside cells can capture molecular machines in action within their native context. In this work, we developed an integrative in-cell structural approach using the ...Structural biology studies performed inside cells can capture molecular machines in action within their native context. In this work, we developed an integrative in-cell structural approach using the genome-reduced human pathogen We combined whole-cell cross-linking mass spectrometry, cellular cryo-electron tomography, and integrative modeling to determine an in-cell architecture of a transcribing and translating expressome at subnanometer resolution. The expressome comprises RNA polymerase (RNAP), the ribosome, and the transcription elongation factors NusG and NusA. We pinpointed NusA at the interface between a NusG-bound elongating RNAP and the ribosome and propose that it can mediate transcription-translation coupling. Translation inhibition dissociated the expressome, whereas transcription inhibition stalled and rearranged it. Thus, the active expressome architecture requires both translation and transcription elongation within the cell.
External linksScience / PubMed:32732422 / PubMed Central
MethodsEM (subtomogram averaging)
Resolution5.6 - 9.7 Å
Structure data

EMDB-10677:
in-cell 70S ribosome from Mycoplasma pneumoniae
Method: EM (subtomogram averaging) / Resolution: 5.6 Å

EMDB-10678:
in-cell elongating expressome from Mycoplasma pneumoniae with focused refinement on RNAP
Method: EM (subtomogram averaging) / Resolution: 9.2 Å

EMDB-10679:
in-cell elongating expressome from Mycoplasma pneumoniae
Method: EM (subtomogram averaging) / Resolution: 8.5 Å

EMDB-10680:
Multibody refinement of RNA polymerase-NusA body of Mycoplasma pneumoniae in-cell expressome
Method: EM (subtomogram averaging) / Resolution: 9.7 Å

EMDB-10681:
Multibody refinement of 30S ribosome body of Mycoplasma pneumoniae in-cell expressome
Method: EM (subtomogram averaging) / Resolution: 8.0 Å

EMDB-10682:
Multibody refinement of 50S ribosome body of Mycoplasma pneumoniae in-cell expressome
Method: EM (subtomogram averaging) / Resolution: 7.6 Å

EMDB-10683:
70S ribosome from chloramphenicol-treated Mycoplasma pneumoniae cells
Method: EM (subtomogram averaging) / Resolution: 6.5 Å

EMDB-10684:
in-cell stalled expressome from pseudouridimycin-treated Mycoplasma pneumoniae
Method: EM (subtomogram averaging) / Resolution: 7.1 Å

EMDB-10685:
Multibody refinement of RNA polymerase body of pseudouridimycin-stalled Mycoplasma pneumoniae in-cell expressome
Method: EM (subtomogram averaging) / Resolution: 8.1 Å

EMDB-10686:
Multibody refinement of 30S ribosome body of pseudouridimycin-stalled Mycoplasma pneumoniae in-cell expressome
Method: EM (subtomogram averaging) / Resolution: 6.8 Å

EMDB-10687:
Multibody refinement of 50S ribosome body of pseudouridimycin-stalled Mycoplasma pneumoniae in-cell expressome
Method: EM (subtomogram averaging) / Resolution: 6.2 Å

Source
  • Mycoplasma pneumoniae M129 (bacteria)

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