Fluorescent lifetime trajectories of a single fluorophore reveal reaction intermediates during transcription initiation

Maria Sorokina, Hye Ran Koh, Smita Patel, Taekjip Ha

Research output: Contribution to journalArticle

30 Citations (Scopus)

Abstract

(Graph Presented) Single molecule (SM) techniques are relatively new additions to the field of biophysics that allow one to manipulate individual molecules and study their behavior. To make these studies more relevant to what actually happens in the cell, one needs to move beyond the studies of individual molecules in isolation and study many different molecules working in concert. This presents a technical challenge as most SM experiments measure only one observable as a function of time, whereas complex biomolecular systems require multidimensional SM analysis. Förster resonance energy transfer (FRET) is one of the most common single molecule approaches and can report on the real time distance changes. However, FRET requires two fluorophores which will ultimately limit the degree of multiplexing in future SM applications. It will be useful if a single fluorophore can be used to provide equivalent information. In this communication, we show that fluorescence lifetime analysis of a single Cy3 fluorophore attached to the promoter region of the DNA can be used to reveal transient reaction intermediates during transcription initiation by T7 RNA polymerase. This work represents the first demonstration of real-time biochemical reactions observed via single molecule fluorescence lifetime trajectories of immobilized molecules.

Original languageEnglish (US)
Pages (from-to)9630-9631
Number of pages2
JournalJournal of the American Chemical Society
Volume131
Issue number28
DOIs
StatePublished - Jul 22 2009

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Reaction intermediates
Fluorophores
Transcription
Energy Transfer
Trajectories
Molecules
Fluorescence
Biophysics
Genetic Promoter Regions
Energy transfer
DNA
RNA
Multiplexing
Large scale systems
Demonstrations

All Science Journal Classification (ASJC) codes

  • Chemistry(all)
  • Biochemistry
  • Catalysis
  • Colloid and Surface Chemistry

Cite this

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Fluorescent lifetime trajectories of a single fluorophore reveal reaction intermediates during transcription initiation. / Sorokina, Maria; Koh, Hye Ran; Patel, Smita; Ha, Taekjip.

In: Journal of the American Chemical Society, Vol. 131, No. 28, 22.07.2009, p. 9630-9631.

Research output: Contribution to journalArticle

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