Visualization and Purification of Caenorhabditis elegans Germ Granule Proteins Using Proximity Labeling

Hertz, Hannah L.; Price, Ian F.; Tang, Wen

doi:10.21769/bioprotoc.4386

Cited by 2 publications

(2 citation statements)

References 14 publications

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“…Our lab recently employed a proximity-based labeling method in combination with mass spectrometry to define P granule constituents 24 , 25 . Among newly identified components, we have characterized several key regulators including LOTUS domain and IDR ( I ntrinsically D isordered R egions)-containing proteins EGGD-1 and −2 ( E mbryonic and G ermline P G ranule D etached), also referred to as MIP-1 and −2 ( M EG-3 i nteracting p rotein) by Gunsalus’s lab 24 , 26 .…”

Section: Introductionmentioning

confidence: 99%

C. elegans germ granules sculpt both germline and somatic RNAome

Price,

Wagner,

Pastore

et al. 2023

Nat Commun

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Germ granules are membrane-less organelles essential for small RNA biogenesis and germline development. Among the conserved properties of germ granules is their association with the nuclear membrane. Recent studies demonstrated that LOTUS domain proteins, EGGD-1 and EGGD-2 (also known as MIP-1 and MIP-2 respectively), promote the formation of perinuclear germ granules in C. elegans. This finding presents a unique opportunity to evaluate the significance of perinuclear localization of germ granules. Here we show that loss of eggd-1 causes the coalescence of germ granules and formation of abnormal cytoplasmic aggregates. Impairment of perinuclear granules affects certain germline classes of small RNAs including Piwi-interacting RNAs. Transcriptome profiling reveals overexpression of spermatogenic and cuticle-related genes in eggd-1 hermaphrodites. We further demonstrate that disruption of germ granules activates HLH-30-mediated transcriptional program in somatic tissues. Collectively, our findings underscore the essential role of EGGD-1 in germ granule organization and reveal an unexpected germ granule-to-soma communication.

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Section: Introductionmentioning

confidence: 99%

C. elegans germ granules sculpt both germline and somatic RNAome

Price,

Wagner,

Pastore

et al. 2023

Nat Commun

Self Cite

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“…Hertz et al applied TurboID labeling and purification to localize proteins in the P granules of Caenorhabditis elegans embryos, revealing the proteome of worm P granules. This method can be used to study other membranous organelles in multicellular organisms [ 137 ].…”

Section: Applications Of Biotin Ligase-based Proximity Marker Technologymentioning

confidence: 99%

The development of proximity labeling technology and its applications in mammals, plants, and microorganisms

Guo,

et al. 2023

Cell Commun Signal

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Protein‒protein, protein‒RNA, and protein‒DNA interaction networks form the basis of cellular regulation and signal transduction, making it crucial to explore these interaction networks to understand complex biological processes. Traditional methods such as affinity purification and yeast two-hybrid assays have been shown to have limitations, as they can only isolate high-affinity molecular interactions under nonphysiological conditions or in vitro. Moreover, these methods have shortcomings for organelle isolation and protein subcellular localization. To address these issues, proximity labeling techniques have been developed. This technology not only overcomes the limitations of traditional methods but also offers unique advantages in studying protein spatial characteristics and molecular interactions within living cells. Currently, this technique not only is indispensable in research on mammalian nucleoprotein interactions but also provides a reliable approach for studying nonmammalian cells, such as plants, parasites and viruses. Given these advantages, this article provides a detailed introduction to the principles of proximity labeling techniques and the development of labeling enzymes. The focus is on summarizing the recent applications of TurboID and miniTurbo in mammals, plants, and microorganisms.

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Visualization and Purification of Caenorhabditis elegans Germ Granule Proteins Using Proximity Labeling

Cited by 2 publications

References 14 publications

C. elegans germ granules sculpt both germline and somatic RNAome

C. elegans germ granules sculpt both germline and somatic RNAome

The development of proximity labeling technology and its applications in mammals, plants, and microorganisms

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