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In vivo pair correlation analysis of EGFP intranuclear diffusion reveals DNA-dependent molecular flow
- Source :
- Hinde, E; Cardarelli, F; Digman, MA; & Gratton, E. (2010). In vivo pair correlation analysis of EGFP intranuclear diffusion reveals DNA-dependent molecular flow. Proceedings of the National Academy of Sciences of the United States of America, 107(38), 16560-16565. doi: 10.1073/pnas.1006731107. UC Irvine: Retrieved from: http://www.escholarship.org/uc/item/5mw799k1
- Publication Year :
- 2010
-
Abstract
- No methods proposed thus far have the capability to measure overall molecular flow in the nucleus of living cells. Here, we apply the pair correlation function analysis (pCF) to measure molecular anisotropic diffusion in the interphase nucleus of live cells. In the pCF method, we cross-correlate fluctuations at several distances and locations within the nucleus, enabling us to define migration paths and barriers to diffusion. We use monomeric EGFP as a prototypical inert molecule and measure flow in and between different nuclear environments. Our results suggest that there are two disconnect molecular flows throughout the nucleus associated with high and low DNA density regions. We show that different density regions of DNA form a networked channel that allows EGFP to diffuse freely throughout, however with restricted ability to traverse the channel. We also observe rare and sudden bursts of molecules traveling across DNA density regions with characteristic time of ≈300 ms, suggesting intrinsic localized change in chromatin structure. This is a unique in vivo demonstration of the intricate chromatin network showing channel directed diffusion of an inert molecule with high spatial and temporal resolution.
- Subjects :
- Anisotropic diffusion
Fluorescent Dye
Green Fluorescent Proteins
Analytical chemistry
Active Transport, Cell Nucleus
CHO Cells
Biology
Transfection
Benzimidazole
Green Fluorescent Protein
Models, Biological
Facilitated Diffusion
chemistry.chemical_compound
Cricetulus
Free molecular flow
Cricetinae
Chromatin organization
medicine
Animals
Fluctuation spectroscopy
Diffusion (business)
Cell Nucleu
Fluorescent Dyes
Cell Nucleus
Multidisciplinary
Microscopy, Confocal
Facilitated diffusion
Active Transport, Cell Nucleu
Animal
Medicine (all)
DNA
Recombinant Protein
Biological Sciences
Nucleus structure
Chromatin
Recombinant Proteins
Cell nucleus
medicine.anatomical_structure
CHO Cell
chemistry
Biophysics
Benzimidazoles
Cricetulu
Nucleus
Subjects
Details
- ISSN :
- 10916490
- Volume :
- 107
- Issue :
- 38
- Database :
- OpenAIRE
- Journal :
- Proceedings of the National Academy of Sciences of the United States of America
- Accession number :
- edsair.doi.dedup.....5b09944156208e21029e0853a17190cb