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Single-molecule narrow-field microscopy of protein-DNA binding dynamics in glucose signal transduction of live yeast cells

Lookup NU author(s): Dr Adam WollmanORCiD


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© Springer Science+Business Media New York 2016. Single-molecule narrow-field microscopy is a versatile tool to investigate a diverse range of protein dynamics in live cells and has been extensively used in bacteria. Here, we describe how these methods can be extended to larger eukaryotic, yeast cells, which contain subcellular compartments. We describe how to obtain single-molecule microscopy data but also how to analyze these data to track and obtain the stoichiometry of molecular complexes diffusing in the cell. We chose glucose mediated signal transduction of live yeast cells as the system to demonstrate these single-molecule techniques as transcriptional regulation is fundamentally a single-molecule problem—a single repressor protein binding a single binding site in the genome can dramatically alter behavior at the whole cell and population level.

Publication metadata

Author(s): Wollman AJM, Leake MC

Editor(s): Mark C. Leake

Publication type: Book Chapter

Publication status: Published

Book Title: Chromosome Architecture: Methods and Protocols

Year: 2016

Volume: 1431

Pages: 5-15

Online publication date: 10/06/2016

Acceptance date: 02/04/2016

Series Title: Methods in Molecular Biology

Publisher: Humana Press Inc.

Place Published: New York


DOI: 10.1007/978-1-4939-3631-1_2

PubMed id: 27283298

Library holdings: Search Newcastle University Library for this item

ISBN: 9781493936311