SM-TRANSCRIPTION
A single-molecule view of initial transcription
| Coordinatore | THE CHANCELLOR, MASTERS AND SCHOLARS OF THE UNIVERSITY OF OXFORD
Organization address
address: University Offices, Wellington Square contact info |
| Nazionalità Coordinatore | United Kingdom [UK] |
| Totale costo | 172˙740 € |
| EC contributo | 172˙740 € |
| Programma | FP7-PEOPLE
Specific programme "People" implementing the Seventh Framework Programme of the European Community for research, technological development and demonstration activities (2007 to 2013) |
| Code Call | FP7-PEOPLE-2009-IEF |
| Funding Scheme | MC-IEF |
| Anno di inizio | 2010 |
| Periodo (anno-mese-giorno) | 2010-03-01 - 2011-08-31 |
Partecipanti
| # | ||||
|---|---|---|---|---|
| 1 |
THE CHANCELLOR, MASTERS AND SCHOLARS OF THE UNIVERSITY OF OXFORD
Organization address
address: University Offices, Wellington Square contact info |
UK (OXFORD) | coordinator | 172˙740.80 |
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Obiettivo del progetto (Objective)
'The overall objective of this project is to study the initial phase of gene-transcription – i.e., the transformation of genetic information from DNA to RNA – at the level of individual molecules. State-of-the-art single-molecule fluorescence spectroscopy will be used in combination with a nanoscale “spectroscopic ruler” (a method based on the phenomenon of fluorescence resonance energy transfer, also known as FRET) to capture the transient intermediates and conformational changes of RNA polymerase, the protein machine that orchestrates transcription. This analysis will complement the static snapshots of transcription complexes, which have been obtained using X-ray crystallography. Single-molecule fluorescence methods are well suited for real-time studies of initial transcription, since the available temporal resolution is sufficient for monitoring conformational changes during the addition of a single nucleotide to an RNA chain. The specific aims are to develop novel real-time assays for detecting promoter-DNA opening and promoter-escape in vitro; to use the in vitro assays for studying the effect of specific DNA sequences on promoter-proximal pausing and promoter escape; and to take the first steps towards developing similar single-molecule FRET assays that report on transcription in living bacterial cells. The proposed work will contribute to the understanding of initial transcription, providing insights applicable to the transcription systems of higher organisms such as humans. The proposed toolbox will also find extensive use in the study of other important protein-DNA interactions, both in vitro and in vivo.'