Optical Methods to Study Protein-DNA Interactions in Vitro and in Living Cells at the Single-Molecule Level

International Journal of Molecular Sciences

Carina Monico¹  Marco Capitanio¹  Gionata Belcastro¹  Francesco Vanzi¹ 

[1] European Laboratory for Non-linear Spectroscopy, Via Nello Carrara 1, 50019 Sesto Fiorentino (FI), Italy; E-Mails:

关键词: single-molecule techniques;    protein-DNA interactions;    facilitated diffusion;    tethered particle motion;    optical tweezers;    magnetic tweezers;    DNA curtains;    single-molecule imaging;    combining single-molecule fluorescence and optical trapping;    in vivo transcription factors dynamics;   

DOI  :  10.3390/ijms14023961

来源: mdpi

【 摘 要 】

The maintenance of intact genetic information, as well as the deployment of transcription for specific sets of genes, critically rely on a family of proteins interacting with DNA and recognizing specific sequences or features. The mechanisms by which these proteins search for target DNA are the subject of intense investigations employing a variety of methods in biology. A large interest in these processes stems from the faster-than-diffusion association rates, explained in current models by a combination of 3D and 1D diffusion. Here, we present a review of the single-molecule approaches at the forefront of the study of protein-DNA interaction dynamics and target search in vitro and in vivo. Flow stretch, optical and magnetic manipulation, single fluorophore detection and localization as well as combinations of different methods are described and the results obtained with these techniques are discussed in the framework of the current facilitated diffusion model.

【 授权许可】

CC BY   
© 2013 by the authors; licensee Molecular Diversity Preservation International, Basel, Switzerland.