In Situ Monitoring of Transiently Formed Molecular Chaperone Assemblies in Bacteria, Yeast, and Human Cells.

Niels Alberts, Yasith Mathangasinghe, Nadinath B Nillegoda*

*Corresponding author for this work

    Research output: Contribution to journalArticleAcademicpeer-review

    Abstract

    J-domain proteins (JDPs) form the largest and the most diverse co-chaperone family in eukaryotic cells. Recent findings show that specific members of the JDP family could form transient heterocomplexes in eukaryotes to fine-tune substrate selection for the 70 kDa heat shock protein (Hsp70) chaperone-based protein disaggregases. The JDP complexes target acute/chronic stress induced aggregated proteins and presumably help assemble the disaggregases by recruiting multiple Hsp70s to the surface of protein aggregates. The extent of the protein quality control (PQC) network formed by these physically interacting JDPs remains largely uncharacterized in vivo. Here, we describe a microscopy-based in situ protein interaction assay named the proximity ligation assay (PLA), which is able to robustly capture these transiently formed chaperone complexes in distinct cellular compartments of eukaryotic cells. Our work expands the employment of PLA from human cells to yeast (Saccharomyces cerevisiae) and bacteria (Escherichia coli), thus rendering an important tool to monitor the dynamics of transiently formed protein assemblies in both prokaryotic and eukaryotic cells.
    Original languageEnglish
    Article numbere60172
    JournalJournal of Visualized Experiments
    Issue number151
    DOIs
    Publication statusPublished - 2-Sep-2019

    Keywords

    • Biology
    • Issue 151
    • Proximity Ligation Assay
    • Chaperone
    • J-domain protein
    • Hsp70
    • human
    • bacteria
    • yeast
    • E. coli
    • S. cerevisiae
    • protein interaction
    • proteostatis
    • ENERGY-TRANSFER FRET
    • STATIONARY-PHASE
    • CO-CHAPERONE
    • J-PROTEINS
    • PROXIMITY
    • LOCALIZATION
    • MICROSCOPY

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