Monitoring condensate dynamics in S. cerevisiae using fluorescence recovery after photobleaching

Macy L Sprunger; Meredith E Jackrel

doi:10.1016/j.xpro.2022.101592

Monitoring condensate dynamics in S. cerevisiae using fluorescence recovery after photobleaching

STAR Protoc. 2022 Jul 31;3(3):101592. doi: 10.1016/j.xpro.2022.101592. eCollection 2022 Sep 16.

Authors

Macy L Sprunger¹, Meredith E Jackrel²

Affiliations

¹ Department of Chemistry, Washington University, St. Louis, MO 63130, USA.
² Department of Chemistry, Washington University, St. Louis, MO 63130, USA. Electronic address: mjackrel@wustl.edu.

Abstract

This protocol describes the use of fluorescence recovery after photobleaching (FRAP) to investigate the dynamics of Matrin-3 (MATR3) condensates in live budding yeast. We detail how to generate yeast strains containing MATR3 with an enhanced green fluorescent protein (eGFP) tag and induce MATR3-eGFP expression. We provide steps to prepare slides of immobilized yeast cells and perform FRAP imaging and data analysis. This protocol can be broadly applied to study condensate dynamics of a range of proteins in different model systems. For complete details on the use and execution of this protocol, please refer to Sprunger et al. (2022).

Keywords: Biophysics; Cell Biology; Microscopy; Model Organisms; Protein Biochemistry.

Monitoring condensate dynamics in S. cerevisiae using fluorescence recovery after photobleaching

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Abstract

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