Poly(dA:dT) Tracts Differentially Modulate Nucleosome Remodeling Activity of RSC and ISW1a Complexes, Exerting Tract Orientation-Dependent and -Independent Effects
Abstract
:1. Introduction
2. Results
2.1. Poly(dA:dT) Tracts Exert a Differential Effect on the Activity of ISW1a and RSC Complexes
2.2. Poly(dA:dT) Tracts Exert Stronger Enhancement of RSC Nucleosome Disassembling Activity When the Nucleosome Core Is Located 5′ of PolyA
2.3. In a Nucleosome Harboring Linker DNA on Both Sides of the Core but a Poly(dA:dT) Tract on Only One Linker, RSC Preferentially Selects This Linker as Exit DNA for Sliding
2.4. A Design That Forces RSC to Mobilize a Tract across the Nucleosome Core Results in Blocking of Its Sliding Activity by One Tract Orientation
2.5. Poly(dA:dT) Tracts Globally Influence the Directionality of ISW1a Sliding Activity
3. Discussion
3.1. Poly(dA:dT) Tracts Exert Orientation-Dependent and -Independent Effects on RSC Activity
3.2. Poly(dA:dT) Tracts Hinder ISW1a Activity When Located in the Way of Histone Octamer Mobilization
3.3. Effects of Poly(dA:dT) Tracts on ISW1a and RSC at NFRs
4. Materials and Methods
4.1. Protein Complexes
4.2. Plasmids, Probes and Nucleosome Reconstitution
4.3. Competitive Nucleosome Reconstitution
4.4. Binding Assays
4.5. Nucleosome Remodeling Assays
4.6. Determination of Nucleosome Sliding Directionality
4.7. Bioinformatics Analyses
Supplementary Materials
Author Contributions
Funding
Institutional Review Board Statement
Informed Consent Statement
Data Availability Statement
Conflicts of Interest
References
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Amigo, R.; Raiqueo, F.; Tarifeño, E.; Farkas, C.; Gutiérrez, J.L. Poly(dA:dT) Tracts Differentially Modulate Nucleosome Remodeling Activity of RSC and ISW1a Complexes, Exerting Tract Orientation-Dependent and -Independent Effects. Int. J. Mol. Sci. 2023, 24, 15245. https://doi.org/10.3390/ijms242015245
Amigo R, Raiqueo F, Tarifeño E, Farkas C, Gutiérrez JL. Poly(dA:dT) Tracts Differentially Modulate Nucleosome Remodeling Activity of RSC and ISW1a Complexes, Exerting Tract Orientation-Dependent and -Independent Effects. International Journal of Molecular Sciences. 2023; 24(20):15245. https://doi.org/10.3390/ijms242015245
Chicago/Turabian StyleAmigo, Roberto, Fernanda Raiqueo, Estefanía Tarifeño, Carlos Farkas, and José L. Gutiérrez. 2023. "Poly(dA:dT) Tracts Differentially Modulate Nucleosome Remodeling Activity of RSC and ISW1a Complexes, Exerting Tract Orientation-Dependent and -Independent Effects" International Journal of Molecular Sciences 24, no. 20: 15245. https://doi.org/10.3390/ijms242015245