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Lack of evidence for condensin or cohesin sequestration on lipid droplets with packing defects

Lipid droplets (LD) are organelles born from the endoplasmic reticulum that store fats and sterols in an apolar manner both as an energy reservoir and for protective purposes. The LD is delimited by a phospholipid monolayer covered by a rich proteome that dynamically evolves depending on the nutriti...

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Detalles Bibliográficos
Autores principales: Mura, Anaïs, Moriel-Carretero, María
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Caltech Library 2021
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC8569452/
https://www.ncbi.nlm.nih.gov/pubmed/34746685
http://dx.doi.org/10.17912/micropub.biology.000497
Descripción
Sumario:Lipid droplets (LD) are organelles born from the endoplasmic reticulum that store fats and sterols in an apolar manner both as an energy reservoir and for protective purposes. The LD is delimited by a phospholipid monolayer covered by a rich proteome that dynamically evolves depending on the nutritional, genetic, pharmacological and environmental cues. Some of these contexts lead to discontinuities in the phospholipid monolayer, termed “packing defects”, that expose LD hydrophobic contents to the surrounding water environment. This triggers the unscheduled binding of proteins with affinity for hydrophobic surfaces, a thermodynamically favorable reaction. We have raised in the past the concern that this titration includes proteins with important roles in the nucleus, which entails a risk of genome instability. Analysis of previously published LD proteomes isolated from cells lacking the transcription factor Ino2p, a prototype of LD bearing packing defects, made us concentrate on two subunits of the cohesin (Smc1p and Smc3p) and one of the condensin (Smc2p) complexes, both essential to promote genome integrity by structuring chromosomes. We report that, in disagreement with the proteomic data, we find no evidence of titration of condensin or cohesin subunits onto LD in ino2∆ cells. Importantly, during our analysis to label LD, we discovered that the addition of the widely used vital dye AUTODOT(TM), which emits in the blue range of the spectrum, leads, specifically in ino2∆, to the artefactual emission of signals in the green channel. We therefore take the opportunity to warn the community of this undesirable aspect when using this dye.