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Prof. Fernando Moreno-Herrero, Universidad Autónoma de Madrid, Spain: Single-molecule approaches to study protein machines involved in DNA metabolism

Speaker Series. Abstract: DNA metabolism is essential for the proper functioning of the cell. It encompasses processes that preserve the stability, structure, and integrity of the genome, including DNA repair mechanisms and higher-order DNA organization. These tasks are performed by complex, specialized protein machineries composed of low-copy number components that operate in a coordinated and often sequential manner. Many of these processes are inherently mechanical, proceeding through a series of events that are exquisitely regulated by force. Traditional biochemical methods provide ensemble-averaged measurements, lack single-molecule resolution, and offer limited access to physical parameters such as force and directionality. As a result, they can obscure rare or atypical molecular behaviors that may be crucial for proper cellular function. In this talk, I will introduce two single-molecule techniques – Magnetic Tweezers and Optical Tweezers – that enable us to study protein machines involved in DNA processing one molecule at a time. Moreover, by combining these approaches with fluorescence microscopy, we can correlate the biological activity of molecular motors with their spatial localization along the DNA. I will highlight several ongoing projects, including investigations of helicases and molecular motors implicated in bacterial and eukaryotic double-strand break repair, as well as studies of ParB and ParB-like proteins involved in bacterial chromosome segregation and gene regulation of multi-drug resistance plasmids.

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