Abstract
The analysis of protein relocalization by fluorescence microscopy has been important for studying processes involved in genome integrity maintenance at the cellular level. Structure-specific endonucleases are required for genome stability, and work in budding yeast has revealed that these proteins accumulate and colocalize at discrete subnuclear foci following DNA damage. Here we describe protocols for fluorescence microscopy analysis of live budding-yeast cells containing fluorescent-tagged proteins that have been useful for the study of endonuclease relocalization during the cell cycle and under DNA-damaging conditions, all of which can be extended to the analysis of other proteins.
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Acknowledgments
Work in J.A. Tercero’s lab is supported by the Spanish Ministry of Science, Innovation and Universities (MCIU) (grant BFU2016-77663-P AEI/FEDER UE). C.P. Lehmann is the recipient of a predoctoral fellowship (FPU16/01137) from MCIU. The CBMSO receives an institutional grant from Fundación Ramón Areces.
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Lehmann, C.P., Saugar, I., Tercero, J.A. (2021). Fluorescence Microscopy for Analysis of Relocalization of Structure-Specific Endonucleases. In: Aguilera, A., Carreira, A. (eds) Homologous Recombination. Methods in Molecular Biology, vol 2153. Humana, New York, NY. https://doi.org/10.1007/978-1-0716-0644-5_35
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DOI: https://doi.org/10.1007/978-1-0716-0644-5_35
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