Structure and Dynamics of Compact Dinucleosomes: Analysis by Electron Microscopy and spFRET
Abstract
:1. Introduction
2. Results
2.1. The Primary Experimental Approach
2.2. Nucleosomal DNA Is Partially Uncoiled in Dinucleosomes
2.3. Multiple Conformations of CODIs Are Present within a Single Band on a Native Gel: spFRET in-Gel (spFRETG) Approach
2.4. Heterogeneity in the Structure of CODIs: Analysis by Electron Microscopy
2.5. Modeling Structure and Dynamics of the Dinucleosome
2.6. DNA Uncoiling Is Differently Affected in Mono- and Dinucleosomes by Histone Chaperone Spt6
3. Discussion
4. Materials and Methods
4.1. Proteins and Nucleosomes
4.2. spFRET Measurements in Solution
4.3. FRET and spFRET Measurements in Gel
4.4. Electron Microscopy
4.5. Incubation of Nucleosomes in the Presence of Histone Chaperones
4.6. Modeling Dinucleosome Structures
Supplementary Materials
Author Contributions
Funding
Institutional Review Board Statement
Informed Consent Statement
Data Availability Statement
Conflicts of Interest
References
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Stefanova, M.E.; Volokh, O.I.; Chertkov, O.V.; Armeev, G.A.; Shaytan, A.K.; Feofanov, A.V.; Kirpichnikov, M.P.; Sokolova, O.S.; Studitsky, V.M. Structure and Dynamics of Compact Dinucleosomes: Analysis by Electron Microscopy and spFRET. Int. J. Mol. Sci. 2023, 24, 12127. https://doi.org/10.3390/ijms241512127
Stefanova ME, Volokh OI, Chertkov OV, Armeev GA, Shaytan AK, Feofanov AV, Kirpichnikov MP, Sokolova OS, Studitsky VM. Structure and Dynamics of Compact Dinucleosomes: Analysis by Electron Microscopy and spFRET. International Journal of Molecular Sciences. 2023; 24(15):12127. https://doi.org/10.3390/ijms241512127
Chicago/Turabian StyleStefanova, Maria E., Olesya I. Volokh, Oleg V. Chertkov, Grigory A. Armeev, Alexey K. Shaytan, Alexey V. Feofanov, Mikhail P. Kirpichnikov, Olga S. Sokolova, and Vasily M. Studitsky. 2023. "Structure and Dynamics of Compact Dinucleosomes: Analysis by Electron Microscopy and spFRET" International Journal of Molecular Sciences 24, no. 15: 12127. https://doi.org/10.3390/ijms241512127