Mechanics of protein-DNA interaction studied with ultra-fast optical tweezers

Carina Monico; Alessia Tempestini; Francesco Vanzi; Francesco S. Pavone; Marco Capitanio

doi:10.1117/12.2052545

8 May 2014 Mechanics of protein-DNA interaction studied with ultra-fast optical tweezers

Carina Monico, Alessia Tempestini, Francesco Vanzi, Francesco S. Pavone, Marco Capitanio

Proceedings Volume 9129, Biophotonics: Photonic Solutions for Better Health Care IV; 91291X (2014) https://doi.org/10.1117/12.2052545
Event: SPIE Photonics Europe, 2014, Brussels, Belgium

Abstract

The lac operon is a well known example of gene expression regulation, based on the specific interaction of Lac repressor protein (LacI) with its target DNA sequence (operator). LacI and other DNA-binding proteins bind their specific target sequences with rates higher than allowed by 3D diffusion alone. Generally accepted models predict a combination of free 3D diffusion and 1D sliding along non-specific DNA. We recently developed an ultrafast force-clamp laser trap technique capable of probing molecular interactions with sub-ms temporal resolution, under controlled pN-range forces. With this technique, we tested the interaction of LacI with two different DNA constructs: a construct with two copies of the O1 operator separated by 300 bp and a construct containing the native E.coli operator sequences. Our measurements show at least two classes of LacI-DNA interactions: long (in the tens of s range) and short (tens of ms). Based on position along the DNA sequence, the observed interactions can be interpreted as specific binding to operator sequences (long events) and transient interactions with nonspecific sequences (short events). Moreover, we observe continuous sliding of the protein along DNA, passively driven by the force applied with the optical tweezers.

Citation Download Citation

Carina Monico, Alessia Tempestini, Francesco Vanzi, Francesco S. Pavone, and Marco Capitanio "Mechanics of protein-DNA interaction studied with ultra-fast optical tweezers", Proc. SPIE 9129, Biophotonics: Photonic Solutions for Better Health Care IV, 91291X (8 May 2014); https://doi.org/10.1117/12.2052545

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