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3D protein dynamics in the cell nucleus
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Singh, Anand P., Galland, Rémi, Finch-Edmondson, Megan L., Grenci, Gianluca, Sibarita, Jean-Baptiste, Studer, Vincent, Viasnoff, Virgile and Saunders, Timothy E. (2017) 3D protein dynamics in the cell nucleus. Biophysical Journal, 112 (1). pp. 133-142. doi:10.1016/j.bpj.2016.11.3196 ISSN 0006-3495.
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Official URL: http://dx.doi.org/10.1016/j.bpj.2016.11.3196
Abstract
The three-dimensional (3D) architecture of the cell nucleus plays an important role in protein dynamics and in regulating gene expression. However, protein dynamics within the 3D nucleus are poorly understood. Here, we present, to our knowledge, a novel combination of 1) single-objective based light-sheet microscopy, 2) photoconvertible proteins, and 3) fluorescence correlation microscopy, to quantitatively measure 3D protein dynamics in the nucleus. We are able to acquire >3400 autocorrelation functions at multiple spatial positions within a nucleus, without significant photobleaching, allowing us to make reliable estimates of diffusion dynamics. Using this tool, we demonstrate spatial heterogeneity in Polymerase II dynamics in live U2OS cells. Further, we provide detailed measurements of human-Yes-associated protein diffusion dynamics in a human gastric cancer epithelial cell line.
Item Type: | Journal Article | ||||||
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Divisions: | Faculty of Science, Engineering and Medicine > Medicine > Warwick Medical School > Biomedical Sciences Faculty of Science, Engineering and Medicine > Medicine > Warwick Medical School |
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Journal or Publication Title: | Biophysical Journal | ||||||
Publisher: | Biophysical Society | ||||||
ISSN: | 0006-3495 | ||||||
Official Date: | 10 January 2017 | ||||||
Dates: |
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Volume: | 112 | ||||||
Number: | 1 | ||||||
Page Range: | pp. 133-142 | ||||||
DOI: | 10.1016/j.bpj.2016.11.3196 | ||||||
Status: | Peer Reviewed | ||||||
Publication Status: | Published | ||||||
Access rights to Published version: | Open Access (Creative Commons) |
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