Structure of a bacterial RNA polymerase holoenzyme open promoter complex
Initiation of transcription is a primary means for controlling gene expression. In bacteria, the RNA polymerase (RNAP) holoenzyme binds and unwinds promoter DNA, forming the transcription bubble of the open promoter complex (RPo). We have determined crystal structures, refined to 4.14 Å-resolution,...
| Main Authors: | , , , , |
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| Format: | Article |
| Language: | English |
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eLife Sciences Publications Ltd
2015-09-01
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| Series: | eLife |
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| Online Access: | https://elifesciences.org/articles/08504 |
| _version_ | 1797997466144997376 |
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| author | Brian Bae Andrey Feklistov Agnieszka Lass-Napiorkowska Robert Landick Seth A Darst |
| author_facet | Brian Bae Andrey Feklistov Agnieszka Lass-Napiorkowska Robert Landick Seth A Darst |
| author_sort | Brian Bae |
| collection | DOAJ |
| description | Initiation of transcription is a primary means for controlling gene expression. In bacteria, the RNA polymerase (RNAP) holoenzyme binds and unwinds promoter DNA, forming the transcription bubble of the open promoter complex (RPo). We have determined crystal structures, refined to 4.14 Å-resolution, of RPo containing Thermus aquaticus RNAP holoenzyme and promoter DNA that includes the full transcription bubble. The structures, combined with biochemical analyses, reveal key features supporting the formation and maintenance of the double-strand/single-strand DNA junction at the upstream edge of the −10 element where bubble formation initiates. The results also reveal RNAP interactions with duplex DNA just upstream of the −10 element and potential protein/DNA interactions that direct the DNA template strand into the RNAP active site. Addition of an RNA primer to yield a 4 base-pair post-translocated RNA:DNA hybrid mimics an initially transcribing complex at the point where steric clash initiates abortive initiation and σA dissociation. |
| first_indexed | 2024-04-11T10:33:32Z |
| format | Article |
| id | doaj.art-4268cf4aea50443fa856258e8b09015b |
| institution | Directory Open Access Journal |
| issn | 2050-084X |
| language | English |
| last_indexed | 2024-04-11T10:33:32Z |
| publishDate | 2015-09-01 |
| publisher | eLife Sciences Publications Ltd |
| record_format | Article |
| series | eLife |
| spelling | doaj.art-4268cf4aea50443fa856258e8b09015b2022-12-22T04:29:21ZengeLife Sciences Publications LtdeLife2050-084X2015-09-01410.7554/eLife.08504Structure of a bacterial RNA polymerase holoenzyme open promoter complexBrian Bae0Andrey Feklistov1Agnieszka Lass-Napiorkowska2Robert Landick3Seth A Darst4Laboratory for Molecular Biophysics, The Rockefeller University, New York, United StatesLaboratory for Molecular Biophysics, The Rockefeller University, New York, United StatesEdward A. Doisy Department of Biochemistry and Molecular Biology, Saint Louis University School of Medicine, St Louis, United StatesDepartment of Biochemistry, University of Wisconsin-madison, Madison, United States; Department of Bacteriology, University of Wisconsin-Madison, Madison, United StatesLaboratory for Molecular Biophysics, The Rockefeller University, New York, United StatesInitiation of transcription is a primary means for controlling gene expression. In bacteria, the RNA polymerase (RNAP) holoenzyme binds and unwinds promoter DNA, forming the transcription bubble of the open promoter complex (RPo). We have determined crystal structures, refined to 4.14 Å-resolution, of RPo containing Thermus aquaticus RNAP holoenzyme and promoter DNA that includes the full transcription bubble. The structures, combined with biochemical analyses, reveal key features supporting the formation and maintenance of the double-strand/single-strand DNA junction at the upstream edge of the −10 element where bubble formation initiates. The results also reveal RNAP interactions with duplex DNA just upstream of the −10 element and potential protein/DNA interactions that direct the DNA template strand into the RNAP active site. Addition of an RNA primer to yield a 4 base-pair post-translocated RNA:DNA hybrid mimics an initially transcribing complex at the point where steric clash initiates abortive initiation and σA dissociation.https://elifesciences.org/articles/08504open promoter compexRNA polymerasetranscriptionX-ray crystallographyThermus aquaticus |
| spellingShingle | Brian Bae Andrey Feklistov Agnieszka Lass-Napiorkowska Robert Landick Seth A Darst Structure of a bacterial RNA polymerase holoenzyme open promoter complex eLife open promoter compex RNA polymerase transcription X-ray crystallography Thermus aquaticus |
| title | Structure of a bacterial RNA polymerase holoenzyme open promoter complex |
| title_full | Structure of a bacterial RNA polymerase holoenzyme open promoter complex |
| title_fullStr | Structure of a bacterial RNA polymerase holoenzyme open promoter complex |
| title_full_unstemmed | Structure of a bacterial RNA polymerase holoenzyme open promoter complex |
| title_short | Structure of a bacterial RNA polymerase holoenzyme open promoter complex |
| title_sort | structure of a bacterial rna polymerase holoenzyme open promoter complex |
| topic | open promoter compex RNA polymerase transcription X-ray crystallography Thermus aquaticus |
| url | https://elifesciences.org/articles/08504 |
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