DC Field | Value | Language |
dc.contributor.author | Zhang, Zhengjian | |
dc.contributor.author | English, Brian P. | |
dc.contributor.author | Grimm, Jonathan B. | |
dc.contributor.author | Kazane, Stephanie A. | |
dc.contributor.author | Hu, Wenxin | |
dc.contributor.author | Tsai, Albert | |
dc.contributor.author | Inouye, Carla | |
dc.contributor.author | You, Changjiang | |
dc.contributor.author | Piehler, Jacob | |
dc.contributor.author | Schultz, Peter G. | |
dc.contributor.author | Lavis, Luke D. | |
dc.contributor.author | Revyakin, Andrey | |
dc.contributor.author | Tjian, Robert | |
dc.date.accessioned | 2021-12-23T16:22:49Z | - |
dc.date.available | 2021-12-23T16:22:49Z | - |
dc.date.issued | 2016 | |
dc.identifier.issn | 08909369 | |
dc.identifier.uri | https://osnascholar.ub.uni-osnabrueck.de/handle/unios/14317 | - |
dc.description.abstract | Transcription of protein-encoding genes in eukaryotic cells requires the coordinated action of multiple general transcription factors (GTFs) and RNA polymerase II (Pol II). A ``step-wise'' preinitiation complex (PIC) assembly model has been suggested based on conventional ensemble biochemical measurements, in which protein factors bind stably to the promoter DNA sequentially to build a functional PIC. However, recent dynamic measurements in live cells suggest that transcription factors mostly interact with chromatin DNA rather transiently. To gain a clearer dynamic picture of PIC assembly, we established an integrated in vitro single-molecule transcription platform reconstituted from highly purified human transcription factors and complemented it by live-cell imaging. Here we performed real-time measurements of the hierarchal promoter-specific binding of TFIID, TFIIA, and TFIIB. Surprisingly, we found that while promoter binding of TFIID and TFIIA is stable, promoter binding by TFIIB is highly transient and dynamic (with an average residence time of 1.5 sec). Stable TFIIB-promoter association and progression beyond this apparent PIC assembly checkpoint control occurs only in the presence of Pol II-TFIIF. This transient-to-stable transition of TFIIB-binding dynamics has gone undetected previously and underscores the advantages of single-molecule assays for revealing the dynamic nature of complex biological reactions. | |
dc.description.sponsorship | Damon Runyon Cancer Research Foundation [DRG2220-15]; BBSRCUK Research & Innovation (UKRI)Biotechnology and Biological Sciences Research Council (BBSRC) [BB/L021730/1] Funding Source: UKRI; Biotechnology and Biological Sciences Research CouncilUK Research & Innovation (UKRI)Biotechnology and Biological Sciences Research Council (BBSRC) [BB/L021730/1] Funding Source: researchfish; We thank Nikola Ivica and Stephen Bell for help with sortase-mediated protein labeling, colleagues at janelia Research Campus of the Howard Hughes Medical Institute and members of the Tjian Laboratory at the University of California at Berkeley for critical reading of the manuscript, and Yan Li and Gina Dailey for molecular cloning assistance. A.T. is a Damon Runyon Fellow supported by the Damon Runyon Cancer Research Foundation (DRG2220-15). | |
dc.language.iso | en | |
dc.publisher | COLD SPRING HARBOR LAB PRESS, PUBLICATIONS DEPT | |
dc.relation.ispartof | GENES & DEVELOPMENT | |
dc.subject | BOX | |
dc.subject | Cell Biology | |
dc.subject | CELLS | |
dc.subject | CORE PROMOTER | |
dc.subject | CRYSTAL-STRUCTURE | |
dc.subject | Developmental Biology | |
dc.subject | DNA-BINDING | |
dc.subject | dynamic analysis | |
dc.subject | fluorescence imaging | |
dc.subject | Genetics & Heredity | |
dc.subject | in vitro reconstitution | |
dc.subject | INITIATION | |
dc.subject | preinitiation complex | |
dc.subject | PROTEIN | |
dc.subject | RNA-POLYMERASE-II | |
dc.subject | single-molecule | |
dc.subject | TBP | |
dc.subject | transcription | |
dc.subject | YEAST TFIIA/TBP/DNA COMPLEX | |
dc.title | Rapid dynamics of general transcription factor TFIIB binding during preinitiation complex assembly revealed by single-molecule analysis | |
dc.type | journal article | |
dc.identifier.doi | 10.1101/gad.285395.116 | |
dc.identifier.isi | ISI:000385901400008 | |
dc.description.volume | 30 | |
dc.description.issue | 18 | |
dc.description.startpage | 2106 | |
dc.description.endpage | 2118 | |
dc.contributor.orcid | 0000-0002-7839-6397 | |
dc.contributor.orcid | 0000-0002-4037-6294 | |
dc.contributor.orcid | 0000-0002-1643-0780 | |
dc.contributor.researcherid | L-3901-2014 | |
dc.contributor.researcherid | O-8156-2016 | |
dc.identifier.eissn | 15495477 | |
dc.publisher.place | 1 BUNGTOWN RD, COLD SPRING HARBOR, NY 11724 USA | |
dcterms.isPartOf.abbreviation | Genes Dev. | |
dcterms.oaStatus | Green Published, gold | |
crisitem.author.dept | Sonderforschungsbereich 944: Physiologie und Dynamik zellulärer Mikrokompartimente | - |
crisitem.author.dept | FB 05 - Biologie/Chemie | - |
crisitem.author.deptid | organisation19 | - |
crisitem.author.deptid | fb05 | - |
crisitem.author.orcid | 0000-0002-7839-6397 | - |
crisitem.author.orcid | 0000-0002-2143-2270 | - |
crisitem.author.parentorg | FB 05 - Biologie/Chemie | - |
crisitem.author.parentorg | Universität Osnabrück | - |
crisitem.author.grandparentorg | Universität Osnabrück | - |
crisitem.author.netid | YoCh745 | - |
crisitem.author.netid | PiJa938 | - |