Blears, Daniel Arthur;
(2023)
Analysis of Integrator assembly and functions in transcription regulation.
Doctoral thesis (Ph.D), UCL (University College London).
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Abstract
Studies of the effects of 5-6-dichloro-1-β-D-ribofuranosylbenzimidazole (DRB) have led to the identification and characterisation of fundamental dynamics in the early stages of the metazoan transcription cycle. These involve a block to transcription shortly after transcription initiation, known as promoter proximal pausing, which is coupled to either pause-release or premature termination of transcription. The metazoan specific Negative elongation factor (NELF) and Integrator complexes are fundamental to the regulation of these dynamics and have been studied extensively since their discovery in the early 2000’s. NELF is crucial for pausing, while premature-termination is mediated by Integrator complex which harbours RNA cleavage and protein phosphatase activities. Many aspects of the mechanism, the composition, assembly, and function of Integrator are unknown. This thesis demonstrates that the INTS6 subunit is critical for the association of a subpopulation of Integrator complexes to chromatin. INTS6-dependent Integrator processes promoter-proximal RNA polymerase II (RNAPII) that contains a novel carboxy-terminal domain (CTD) phosphorylation pattern. Interestingly, loss of INTS6 results in the recruitment of a set of new factors to the paused complex that may serve as essential back-up mechanisms to allow the removal of paused RNAPII and the maintenance of genome stability. Moreover, the uncharacterised INTS12 subunit is shown to be critical for Integrator function. This subunit harbours a highly unique plant homeodomain (PHD) finger domain, which - rather than interacting with Histone H3 tails - directly interacts with a domain in NELFA; hence a novel mode of PHD finger regulation is uncovered. Taken together, this thesis advances a new understanding of Integrator and NELF functions in pausing and premature termination, while providing a new perspective on the biology of the conserved family of PHD finger proteins.
| Type: | Thesis (Doctoral) |
|---|---|
| Qualification: | Ph.D |
| Title: | Analysis of Integrator assembly and functions in transcription regulation |
| Open access status: | An open access version is available from UCL Discovery |
| Language: | English |
| Additional information: | Copyright © The Author 2022. Original content in this thesis is licensed under the terms of the Creative Commons Attribution-NonCommercial 4.0 International (CC BY-NC 4.0) Licence (https://creativecommons.org/licenses/by-nc/4.0/). Any third-party copyright material present remains the property of its respective owner(s) and is licensed under its existing terms. Access may initially be restricted at the author’s request. |
| UCL classification: | UCL UCL > Provost and Vice Provost Offices > School of Life and Medical Sciences UCL > Provost and Vice Provost Offices > School of Life and Medical Sciences > Faculty of Life Sciences UCL > Provost and Vice Provost Offices > School of Life and Medical Sciences > Faculty of Life Sciences > Div of Biosciences |
| URI: | https://discovery.ucl.ac.uk/id/eprint/10174822 |
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