Menu
MapConnectionsTopicsBulgariaThe Night (K-TRIO)Marie Skłodowska-CurieFor studentsFor teachersMethodologySurprise me
H2020Individual fellowship2016–2017

G4invivo · Probing intracellular folding and dynamics of telomeric DNA structures with single-molecule FRET

Horizon 2020 — Marie Skłodowska-Curie Actions

Duration
2016-01-01 → 2017-12-31
EU contribution
€212,195
Participants
1
Scheme
MSCA-IF-EF-ST

Lines connect the coordinator with its partners.

Results in brief

Probing intracellular folding and dynamics of telomeric DNA structures with single-molecule FRET

Non-canonical DNA secondary structures can participate in several biological processes, such as transcription, replication and telomere lengthening, etc. G-quadruplexes are examples of such non-canonical DNA structures that, in particular, are expected to occur at telomeres and gene promoters regions of the human genome. Several recent observations strongly support the occurrence and functional relevance of G-quadruplexes in vivo, however a detailed description of their behaviour under cellular conditions is currently missing. Their investigation under physiological settings is important in order to understand the mechanisms of their action at the cellular level and furthermore use them as potential drug targets. Telomeric G-quadruplexes, in particular, are polymorphic and dynamical structures that can form a variety of molecular structures under different experimental conditions. This intrinsic polymorphism and dynamics is difficult to resolve with the majority of classical biophysical techniques that provide both structure and time averaged ensemble results. In order to overcome these complications, we used high-resolution single-molecule fluorescence microscopy techniques. These methods allow probing the behavior of G-quadruplex molecules one by one and thus allow obtaining a direct view of both their structure and dynamics. The objective of the project was to probe the conformation and structural dynamics of G-quadruplex DNA under a range of cell-mimicking milieu to obtain a mechanistic understanding of their behavior.

Data: CORDIS, © European Union

Project objective

Telomeres, the ends of linear eukaryotic chromosomes, are one of the most versatile “molecular machines” of life. They perform a vital role in several fundamental processes in biology, as maintenance of genome integrity and the regulation of cell growth. Human telomeric DNA consists of thousands of tandem repeats of guanine-rich sequences, terminating the chromosomes. Under physiological conditions, such guanine-rich repeats are capable of adopting four stranded structures, called G-quadruplex DNA. Considerable research has been dedicated to an understanding of functions of telomeres, and during the past decades, much progress has been made towards identifying the underlying processes for G-quadruplex DNA isolated in vitro. However, most of our knowledge of conformation and mechanistic understanding of the balance between the structure and function of telomeric DNA inside cells has remained very limited, largely due to a lack of suitable methods. An opportunity to bridge this gap is single-molecule fluorescence microscopy in combination with Förster resonance energy transfer (FRET), a powerful technique that allows distances, distance distributions, and dynamics of individual biological molecules to be measured even in complex and heterogeneous environments, as cellular milieu.The goal of this project is to employ such optical single-molecule techniques to perform a comprehensive biophysical analysis of the folding and dynamics of G-quadruplex DNA within live cells. This investigation will provide an idea of how accurately our current understanding of the folding and dynamics of G-quadruplex DNA in vitro describes the situation inside the living cell, and will, among other aspects, be crucial for understanding the biological functions of G-quadruplexes and in particular may stimulate the enhancement of the efficacy of the novel anticancer therapies, which use these structures as an active target for drug development.

Original text from CORDIS.

Participants

  • AARHUS UNIVERSITET · Aarhus CCoordinatorDenmark

Links

Data: CORDIS, © European Union