Obiettivo
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.
Campo scientifico (EuroSciVoc)
CORDIS classifica i progetti con EuroSciVoc, una tassonomia multilingue dei campi scientifici, attraverso un processo semi-automatico basato su tecniche NLP. Cfr.: https://op.europa.eu/en/web/eu-vocabularies/euroscivoc.
CORDIS classifica i progetti con EuroSciVoc, una tassonomia multilingue dei campi scientifici, attraverso un processo semi-automatico basato su tecniche NLP. Cfr.: https://op.europa.eu/en/web/eu-vocabularies/euroscivoc.
- scienze naturaliscienze biologichegeneticaDNA
- scienze naturaliscienze della terra e scienze ambientali connessegeologiamineralogiacristallografia
- scienze naturaliscienze fisicheotticaspettroscopiaspettroscopia a emissione
- scienze naturaliscienze biologichegeneticanucleotidi
- scienze naturaliscienze biologichegeneticaRNA
È necessario effettuare l’accesso o registrarsi per utilizzare questa funzione
Argomento(i)
Invito a presentare proposte
FP7-PEOPLE-2009-IEF
Vedi altri progetti per questo bando
Meccanismo di finanziamento
MC-IEF - Intra-European Fellowships (IEF)Coordinatore
OX1 2JD Oxford
Regno Unito