Dexl, Stefan and Reichelt, Robert and Kraatz, Katharina and Schulz, Sarah and Grohmann, Dina and Bartlett, Michael and Thomm, Michael (2018) Displacement of the transcription factor B reader domain during transcription initiation. NUCLEIC ACIDS RESEARCH, 46 (19). pp. 10066-10081. ISSN 0305-1048, 1362-4962
Full text not available from this repository. (Request a copy)Abstract
Transcription initiation by archaeal RNA polymerase (RNAP) and eukaryotic RNAP II requires the general transcription factor (TF) B/IIB. Structural analyses of eukaryotic transcription initiation complexes locate the B-reader domain of TFIIB in close proximity to the active site of RNAP II. Here, we present the first crosslinking mapping data that describe the dynamic transitions of an archaeal TFB to provide evidence for structural rearrangements within the transcription complex during transition from initiation to early elongation phase of transcription. Using a highly specific UV-inducible crosslinking system based on the unnatural amino acid para-benzoyl-phenylalanine allowed us to analyze contacts of the Pyrococcus furiosus TFB B-reader domain with site-specific radiolabeled DNA templates in preinitiation and initially transcribing complexes. Crosslink reactions at different initiation steps demonstrate interactions of TFB with DNA at registers +6 to +14, and reduced contacts at +15, with structural transitions of the B-reader domain detected at register +10. Our data suggest that the B-reader domain of TFB interacts with nascent RNA at register +6 and +8 and it is displaced from the transcribed-strand during the transition from +9 to +10, followed by the collapse of the transcription bubble and release of TFB from register +15 onwards.
| Item Type: | Article |
|---|---|
| Uncontrolled Keywords: | RNA-POLYMERASE-II; TATA-BINDING PROTEIN; PHOTO-CROSS-LINKING; N-TERMINAL REGION; OPEN COMPLEX; ARCHAEAL TRANSCRIPTION; PREINITIATION COMPLEX; STRUCTURAL BASIS; ABORTIVE INITIATION; PROMOTER CLEARANCE; |
| Subjects: | 500 Science > 570 Life sciences |
| Divisions: | Biology, Preclinical Medicine > Institut für Biochemie, Genetik und Mikrobiologie > Lehrstuhl für Mikrobiologie > Prof. Dr. Dina Grohmann |
| Depositing User: | Dr. Gernot Deinzer |
| Date Deposited: | 10 Oct 2019 06:55 |
| Last Modified: | 10 Oct 2019 06:55 |
| URI: | https://pred.uni-regensburg.de/id/eprint/13558 |
Actions (login required)
 |
View Item |
