| dc.contributor.advisor | Cramer, Patrick Prof. Dr. | |
| dc.contributor.author | Dienemann, Christian | |
| dc.date.accessioned | 2019-02-11T09:36:22Z | |
| dc.date.available | 2019-02-11T09:36:22Z | |
| dc.date.issued | 2019-02-11 | |
| dc.identifier.uri | http://hdl.handle.net/11858/00-1735-0000-002E-E587-B | |
| dc.identifier.uri | http://dx.doi.org/10.53846/goediss-7215 | |
| dc.language.iso | eng | de |
| dc.rights.uri | http://creativecommons.org/licenses/by-nc-nd/4.0/ | |
| dc.subject.ddc | 572 | de |
| dc.subject.ddc | 571.4 | de |
| dc.title | Structural and Functional Investigation of Promoter Distortion and Opening in the RNA Polymerase II Cleft | de |
| dc.type | doctoralThesis | de |
| dc.contributor.referee | Cramer, Patrick Prof. Dr. | |
| dc.date.examination | 2018-04-09 | |
| dc.description.abstracteng | Transcription of genes is a central process of life that defines how genetic information is
used. This determines the metabolic state, morphology and fate of a cell, tissue, organ and
the entire organism. In eukaryotes, the most regulated step of transcription is initiation
during which RNA polymerase II (Pol II) is specifically recruited to the gene promoter by
assembling with transcription factors (TF) –IIA, -IIB, -IID, -IIF, -IIE, -IIH and the co-
activator Mediator complex. For initial transcription, the double helix of the promoter
DNA of the closed complex (CC) has to be opened in the initially melted region (IMR).
The open complex (OC) then provides the single stranded template DNA that can be
transcribed. Promoter opening was shown to require the DNA translocase activity of
TFIIH; however, spontaneous DNA melting could also be observed. The molecular
pathway of TFIIH dependent and spontaneous promoter melting is still unknown due to a
lack of high resolution structural studies targeting intermediates of the CC-to-OC
transition.
Here I report the structure of a new transcription initiation intermediate just before
promoter DNA opening at ~5 Å resolution. This structure reveals closed distorted
promoter DNA in the Pol II active center cleft. The distortion is induced by the closed Pol
II clamp domain and stabilized by structural elements of TFIIF and TFIIE. The distorted
DNA shows a helical axis offset and underwinding of the closed promoter DNA that
weaken the DNA and prime it for melting. I further present in vivo and in vitro data
showing that the DNA duplex stability of the IMR determines whether DNA
spontaneously melts after distortion or whether it remains closed and requires TFIIH
activity to be melted. By systematic comparison of Pol I and Pol III promoters with the Pol
II system, I find that promoters of the other nuclear RNA polymerases also contain a weak
DNA duplex in their IMR. This explains why they do not need a TFIIH homologue or
ATP-hydrolysis to open promoter DNA. This suggests a unified mechanism of promoter
DNA melting that involves DNA distortion by clamp closure and subsequent melting of
the IMR. Taken together, these results greatly improve our understanding of the molecular
mechanism of promoter DNA melting by nuclear RNA polymerases and for the first time
provide a general mechanism for promoter opening. | de |
| dc.contributor.coReferee | Ficner, Ralf Prof. Dr. | |
| dc.subject.eng | transcription | de |
| dc.subject.eng | RNA polymerase II | de |
| dc.subject.eng | TFIIH | de |
| dc.subject.eng | Ssl2 | de |
| dc.subject.eng | XPB | de |
| dc.subject.eng | promoter opening | de |
| dc.subject.eng | DNA distortion | de |
| dc.identifier.urn | urn:nbn:de:gbv:7-11858/00-1735-0000-002E-E587-B-8 | |
| dc.affiliation.institute | Göttinger Graduiertenschule für Neurowissenschaften, Biophysik und molekulare Biowissenschaften (GGNB) | de |
| dc.subject.gokfull | Biologie (PPN619462639) | de |
| dc.identifier.ppn | 1048915077 | |