3D Protein Dynamics in the Cell Nucleus.

Anand P. Singh, Rémi Galland, Megan L. Finch-Edmondson, Gianluca Grenci, Jean-Baptiste Sibarita, Vincent Studer, Virgile Viasnoff, Timothy E. Saunders
Biophysical Journal. 2017-01-01; 112(1): 133-142
DOI: 10.1016/j.bpj.2016.11.3196

PubMed
Read on PubMed



1. Biophys J. 2017 Jan 10;112(1):133-142. doi: 10.1016/j.bpj.2016.11.3196.

3D Protein Dynamics in the Cell Nucleus.

Singh AP(1), Galland R(2), Finch-Edmondson ML(3), Grenci G(1), Sibarita JB(2),
Studer V(2), Viasnoff V(4), Saunders TE(5).

Author information:
(1)Mechanobiology Institute, National University of Singapore, Singapore.
(2)Institut Interdisciplinaire de Neurosciences, University of Bordeaux, France;
Centre National de la Recherche Scientifique UMR 5297, University of Bordeaux,
France.
(3)Mechanobiology Institute, National University of Singapore, Singapore;
Department of Physiology, National University of Singapore, Singapore.
(4)Mechanobiology Institute, National University of Singapore, Singapore;
Department of Biological Sciences, National University of Singapore, Singapore;
Centre National de la Recherche Scientifique UMI 3639, Singapore. Electronic
address: .
(5)Mechanobiology Institute, National University of Singapore, Singapore;
Department of Biological Sciences, National University of Singapore, Singapore;
Institute for Molecular and Cell Biology, A(∗)Star, Singapore. Electronic
address: .

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.

Copyright © 2017 Biophysical Society. Published by Elsevier Inc. All rights
reserved.

DOI: 10.1016/j.bpj.2016.11.3196
PMCID: PMC5232349
PMID: 28076804 [Indexed for MEDLINE]

Know more about