Journal article
Radial pair correlation of molecular brightness fluctuations maps protein diffusion as a function of oligomeric state within live-cell nuclear architecture
A Solano, J Lou, L Scipioni, E Gratton, E Hinde
Biophysical Journal | CELL PRESS | Published : 2022
Abstract
Nuclear proteins can modulate their DNA binding activity and the exploration volume available during DNA target search by self-associating into higher-order oligomers. Directly tracking this process in the nucleoplasm of a living cell is, however, a complex task. Thus, here we present a microscopy method based on radial pair correlation of molecular brightness fluctuations (radial pCOMB) that can extract the mobility of a fluorescently tagged nuclear protein as a function of its oligomeric state and spatiotemporally map the anisotropy of this parameter with respect to nuclear architecture. By simply performing a rapid frame scan acquisition, radial pCOMB has the capacity to detect, within ea..
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Grants
Awarded by National Institutes of Health
Funding Acknowledgements
A.S. and J.L. are supported by an Australian Research Council (ARC) discovery project (DP180101387 and DP210102984) . E.H. is supported by an Australian Research Council Future Fellowship (FT200100401) and the Jacob Haimson Beverly Mecklenburg Lectureship. We thank the Biological Optical Microscopy Platform, University of Melbourne, for enabling access to the Olympus FV3000 confocal laser scanning microscope. We thank A/Prof. Marie Bogoyevitch and Prof. David Jans for providing reagents. We thank Dr. Toan Nguyen from the Monash Immersive Visualisation Plat-form for his input and advice toward development of custom MATLAB code for radial pCOMB analysis. E.G. and L.S are supported by grants from the US National Institutes of Health (P41-GM103540 and P50-GM076516) .