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Protein Aggregation pp 543–560Cite as

An Optimized Stress Granule Detection Method: Investigation of UBQLN2 Effect on Stress Granule Formation

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Part of the book series: Methods in Molecular Biology ((MIMB,volume 2551))

Abstract

Stress granules (SGs) are cytosolic ribonucleoprotein granules that form via a liquid–liquid phase separation in response to environmental stresses such as heat, oxidative, and osmotic changes. Due to the condensation of low complexity, hydrophobic regions in core SG components in these highly dynamic granules, defects in SG maintenance and formation have been linked to toxic aggregate formation in neurodegenerative diseases such as amyotrophic lateral sclerosis and frontotemporal dementia. However, efforts to dissect mechanisms regulating SG formation and maintenance have been limited by methods of tracking protein–SG localization. Here we describe a method for detecting and quantifying recruitment of cytosolically enriched proteins to SGs by indirect immunofluorescence microscopy. Using this method, we tracked the transient recruitment of the cytosolically enriched ubiquitin-like protein, ubiquilin 2 (UBQLN2), and a number of other factors into SGs, demonstrating its utility (Alexander et al., Proc Natl Acad Sci U S A 115:E11485–E11494, 2018).

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Acknowledgments

This work was supported by grants from NIH (NS074324, NS089616, NS110098). We thank the members of Wang lab for discussion.

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Authors and Affiliations

  1. Department of Biochemistry and Molecular Biology, Johns Hopkins University Bloomberg School of Public Health, Baltimore, MD, USA

    Elizabeth J. Alexander & Jiou Wang

  2. Department of Neuroscience, Johns Hopkins University School of Medicine, Baltimore, MD, USA

    Elizabeth J. Alexander & Jiou Wang

Authors
  1. Elizabeth J. Alexander
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  2. Jiou Wang
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Corresponding author

Correspondence to Jiou Wang .

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Editors and Affiliations

  1. Cambridge, MA, USA

    Andrzej Stanisław Cieplak

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Alexander, E.J., Wang, J. (2023). An Optimized Stress Granule Detection Method: Investigation of UBQLN2 Effect on Stress Granule Formation. In: Cieplak, A.S. (eds) Protein Aggregation. Methods in Molecular Biology, vol 2551. Humana, New York, NY. https://doi.org/10.1007/978-1-0716-2597-2_33

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  • DOI: https://doi.org/10.1007/978-1-0716-2597-2_33

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  • Publisher Name: Humana, New York, NY

  • Print ISBN: 978-1-0716-2596-5

  • Online ISBN: 978-1-0716-2597-2

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