Dynamic Architecture of a Minimal RNA Polymerase II Open Promoter Complex
The open promoter complex (OC) is a central intermediate during transcription initiation that contains a DNA bubble. Here, we employ single-molecule Förster resonance energy transfer experiments and Nano-Positioning System analysis to determine the three-dimensional architecture of a minimal OC cons...
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| Veröffentlicht in: | Molecular cell 2012-04, Vol.46 (2), p.136-146 |
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| creator | Treutlein, Barbara Muschielok, Adam Andrecka, Joanna Jawhari, Anass Buchen, Claudia Kostrewa, Dirk Hög, Friederike Cramer, Patrick Michaelis, Jens |
| description | The open promoter complex (OC) is a central intermediate during transcription initiation that contains a DNA bubble. Here, we employ single-molecule Förster resonance energy transfer experiments and Nano-Positioning System analysis to determine the three-dimensional architecture of a minimal OC consisting of promoter DNA, including a TATA box and an 11-nucleotide mismatched region around the transcription start site, TATA box-binding protein (TBP), RNA polymerase (Pol) II, and general transcription factor (TF)IIB and TFIIF. In this minimal OC, TATA-DNA and TBP reside above the Pol II cleft between clamp and protrusion domains. Downstream DNA is dynamically loaded into and unloaded from the Pol II cleft at a timescale of seconds. The TFIIB core domain is displaced from the Pol II wall, where it is located in the closed promoter complex. These results reveal large overall structural changes during the initiation-elongation transition, which are apparently accommodated by the intrinsic flexibility of TFIIB.
► Upstream promoter DNA and TBP above cleft in Pol II open promoter complex ► Major rearrangement of upstream DNA, TBP, and TFIIB during closed-open transition ► Downstream DNA in open complex dynamically switching between two conformations |
| doi_str_mv | 10.1016/j.molcel.2012.02.008 |
| format | Article |
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► Upstream promoter DNA and TBP above cleft in Pol II open promoter complex ► Major rearrangement of upstream DNA, TBP, and TFIIB during closed-open transition ► Downstream DNA in open complex dynamically switching between two conformations</description><identifier>ISSN: 1097-2765</identifier><identifier>EISSN: 1097-4164</identifier><identifier>DOI: 10.1016/j.molcel.2012.02.008</identifier><identifier>PMID: 22424775</identifier><language>eng</language><publisher>United States: Elsevier Inc</publisher><subject>DNA ; DNA-directed RNA polymerase ; energy transfer ; Fluorescence Resonance Energy Transfer ; Models, Genetic ; Promoter Regions, Genetic ; Promoters ; RNA Polymerase II - chemistry ; Saccharomyces cerevisiae - genetics ; Saccharomyces cerevisiae Proteins - chemistry ; TATA box ; TATA-binding protein ; Transcription factors ; Transcription initiation ; Transcription initiation factor TFIIB ; Transcription Initiation Site ; Transcription, Genetic</subject><ispartof>Molecular cell, 2012-04, Vol.46 (2), p.136-146</ispartof><rights>2012 Elsevier Inc.</rights><rights>Copyright © 2012 Elsevier Inc. All rights reserved.</rights><lds50>peer_reviewed</lds50><oa>free_for_read</oa><woscitedreferencessubscribed>false</woscitedreferencessubscribed><citedby>FETCH-LOGICAL-c465t-fcc0f44a4587d828fd3ae9b37fa15b330bf811fd60af764cfccf023680af2ac03</citedby><cites>FETCH-LOGICAL-c465t-fcc0f44a4587d828fd3ae9b37fa15b330bf811fd60af764cfccf023680af2ac03</cites></display><links><openurl>$$Topenurl_article</openurl><openurlfulltext>$$Topenurlfull_article</openurlfulltext><thumbnail>$$Tsyndetics_thumb_exl</thumbnail><linktohtml>$$Uhttps://www.sciencedirect.com/science/article/pii/S1097276512001657$$EHTML$$P50$$Gelsevier$$Hfree_for_read</linktohtml><link.rule.ids>314,776,780,3537,27901,27902,65306</link.rule.ids><backlink>$$Uhttps://www.ncbi.nlm.nih.gov/pubmed/22424775$$D View this record in MEDLINE/PubMed$$Hfree_for_read</backlink></links><search><creatorcontrib>Treutlein, Barbara</creatorcontrib><creatorcontrib>Muschielok, Adam</creatorcontrib><creatorcontrib>Andrecka, Joanna</creatorcontrib><creatorcontrib>Jawhari, Anass</creatorcontrib><creatorcontrib>Buchen, Claudia</creatorcontrib><creatorcontrib>Kostrewa, Dirk</creatorcontrib><creatorcontrib>Hög, Friederike</creatorcontrib><creatorcontrib>Cramer, Patrick</creatorcontrib><creatorcontrib>Michaelis, Jens</creatorcontrib><title>Dynamic Architecture of a Minimal RNA Polymerase II Open Promoter Complex</title><title>Molecular cell</title><addtitle>Mol Cell</addtitle><description>The open promoter complex (OC) is a central intermediate during transcription initiation that contains a DNA bubble. Here, we employ single-molecule Förster resonance energy transfer experiments and Nano-Positioning System analysis to determine the three-dimensional architecture of a minimal OC consisting of promoter DNA, including a TATA box and an 11-nucleotide mismatched region around the transcription start site, TATA box-binding protein (TBP), RNA polymerase (Pol) II, and general transcription factor (TF)IIB and TFIIF. In this minimal OC, TATA-DNA and TBP reside above the Pol II cleft between clamp and protrusion domains. Downstream DNA is dynamically loaded into and unloaded from the Pol II cleft at a timescale of seconds. The TFIIB core domain is displaced from the Pol II wall, where it is located in the closed promoter complex. These results reveal large overall structural changes during the initiation-elongation transition, which are apparently accommodated by the intrinsic flexibility of TFIIB.
► Upstream promoter DNA and TBP above cleft in Pol II open promoter complex ► Major rearrangement of upstream DNA, TBP, and TFIIB during closed-open transition ► Downstream DNA in open complex dynamically switching between two conformations</description><subject>DNA</subject><subject>DNA-directed RNA polymerase</subject><subject>energy transfer</subject><subject>Fluorescence Resonance Energy Transfer</subject><subject>Models, Genetic</subject><subject>Promoter Regions, Genetic</subject><subject>Promoters</subject><subject>RNA Polymerase II - chemistry</subject><subject>Saccharomyces cerevisiae - genetics</subject><subject>Saccharomyces cerevisiae Proteins - chemistry</subject><subject>TATA box</subject><subject>TATA-binding protein</subject><subject>Transcription factors</subject><subject>Transcription initiation</subject><subject>Transcription initiation factor TFIIB</subject><subject>Transcription Initiation Site</subject><subject>Transcription, Genetic</subject><issn>1097-2765</issn><issn>1097-4164</issn><fulltext>true</fulltext><rsrctype>article</rsrctype><creationdate>2012</creationdate><recordtype>article</recordtype><sourceid>EIF</sourceid><recordid>eNqNkUtrGzEQgEVpaBKn_6C0OvZiZ6TVauVLwbh5GPKirc9C1o5amd2VI61D_O8js06OoTAwEnwzmvlEyBcGEwZMnq8nbWgsNhMOjE8gB6gP5ITBtBoLJsXHw5lXsjwmpymtAZgo1fQTOeZccFFV5QlZ_Nx1pvWWzqL953u0_TYiDY4aeus735qG_rqb0YfQ7FqMJiFdLOj9Bjv6EEMbeox0HtpNg89n5MiZJuHnQx6R5eXFn_n1-Ob-ajGf3YytkGU_dtaCE8LkSapaceXqwuB0VVTOsHJVFLByijFXSzCuksJm3gEvpMp3biwUI_J96LuJ4XGLqdetT9lDYzoM26QZcFAlFPx_UAZCTUumMioG1MaQUkSnNzFvH3cZ2nNSr_XgW-99a8gB-7Kvhxe2qxbrt6JXwRn4NgDOBG3-Rp_08nfuIAGAc16ITPwYCMzSnjxGnazHzmLtY_4OXQf__gwvKcWa4w</recordid><startdate>20120427</startdate><enddate>20120427</enddate><creator>Treutlein, Barbara</creator><creator>Muschielok, Adam</creator><creator>Andrecka, Joanna</creator><creator>Jawhari, Anass</creator><creator>Buchen, Claudia</creator><creator>Kostrewa, Dirk</creator><creator>Hög, Friederike</creator><creator>Cramer, Patrick</creator><creator>Michaelis, Jens</creator><general>Elsevier Inc</general><scope>6I.</scope><scope>AAFTH</scope><scope>FBQ</scope><scope>CGR</scope><scope>CUY</scope><scope>CVF</scope><scope>ECM</scope><scope>EIF</scope><scope>NPM</scope><scope>AAYXX</scope><scope>CITATION</scope><scope>7X8</scope><scope>7TM</scope></search><sort><creationdate>20120427</creationdate><title>Dynamic Architecture of a Minimal RNA Polymerase II Open Promoter Complex</title><author>Treutlein, Barbara ; 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Here, we employ single-molecule Förster resonance energy transfer experiments and Nano-Positioning System analysis to determine the three-dimensional architecture of a minimal OC consisting of promoter DNA, including a TATA box and an 11-nucleotide mismatched region around the transcription start site, TATA box-binding protein (TBP), RNA polymerase (Pol) II, and general transcription factor (TF)IIB and TFIIF. In this minimal OC, TATA-DNA and TBP reside above the Pol II cleft between clamp and protrusion domains. Downstream DNA is dynamically loaded into and unloaded from the Pol II cleft at a timescale of seconds. The TFIIB core domain is displaced from the Pol II wall, where it is located in the closed promoter complex. These results reveal large overall structural changes during the initiation-elongation transition, which are apparently accommodated by the intrinsic flexibility of TFIIB.
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| subjects | DNA DNA-directed RNA polymerase energy transfer Fluorescence Resonance Energy Transfer Models, Genetic Promoter Regions, Genetic Promoters RNA Polymerase II - chemistry Saccharomyces cerevisiae - genetics Saccharomyces cerevisiae Proteins - chemistry TATA box TATA-binding protein Transcription factors Transcription initiation Transcription initiation factor TFIIB Transcription Initiation Site Transcription, Genetic |
| title | Dynamic Architecture of a Minimal RNA Polymerase II Open Promoter Complex |
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