Quantifying nucleosome dynamics and protein binding with PIE-FCCS and spFRET

Martens, C.L.G. (2023)

Doctoral Thesis

In this thesis I describe the results of Pulsed Interleaved Excitation and Fluorescence (Cross) Correlation Spectroscopy (PIE-F(C)CS) combined with single-pair Förster Resonance Energy Transfer (spFRET) used to study dynamics in single nucleosomes, which depends on subtle differences in the length of DNA ends, DNA sequence, histone variants and specific and non-specific protein interactions. This technique, which can resolve distances between two fluorophores of only a few nanometers, is an excellent technique to monitor changes in nucleosomal compaction, as the nucleosome is only ten nanometers in diameter. In combination with F(C)CS and PIE, spFRET makes it possible to monitor conformational dynamics on a timescale of micro- to milliseconds.

• Fluorescence Correlation Spectroscopy
• Single pair FRET
• Chromatin
• Nucleosome dynamics
• DNA-protein interactions

• All authors: Martens, C.L.G.
• Supervisor: Noort, S.J.T. van
• Committee: Joo, C.; Schiessel, H.; Aarts, J.; Huber, M.I.; Schaaf, M.J.M.; Schmidt, T.
• Qualification: Doctor (dr.)
• Awarding Institution: Leiden Institute of Physics (LION), Faculty of Science, Leiden University
• Date: 2023-02-01
• Title of host publication: Casimir PhD Series
• ISBN (print): 9789085935506
• Name: 2023-02