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Biology subjects

Siketanc, M.

Publications and source records attributed to Siketanc, M..

2 recordsLinked to original sources

Non-translated mRNA levels determine P-body properties

Translational repression enables rapid adaptation to environmental changes. Under stress, translational repressed mRNA and mRNA decay factors accumulate in cytoplasmic processing bodies (PBs), implicated in mRNA storage and decay. PBs have been mostly studied under glucose starvation in yeast, yet, knowledge is limited under other stress conditions. Here, we identify a correlation between the level of translation attenuation and the number, brightness, fluidity and recruitment of PB core components. Stresses triggering strong translation attenuation caused the formation of few bright and more fluid PBs that recruit the decay factors en bloc. Conversely, weaker translation attenuation induced numerous, dim, more viscous PBs to which PB proteins were sequentially recruited. Importantly, increasing non-translated mRNA levels augmented the brightness of dim PBs and accelerated decay machinery recruitment. Finally, boosting RNA levels increased the size of Dhh1 helicase-containing droplets in vitro. Taken together, we propose a model in which the assembly pathway and biophysical properties of PBs are governed by non-translated mRNA abundance. TeaserBiophysical properties, protein composition and assembly pathways of processing bodies are dependent on available mRNA levels.

cell biology↗

Tag with Caution - How protein tagging influences the formation of condensates

Fluorescent proteins and peptide tags are essential tools in cellular biology, but can alter the biochemical properties of target proteins. Biomolecular condensates, which have emerged as key principles of cellular organization, are suggested to provide robustness to cells, yet they can also respond sensitively to small changes in environmental conditions--or tagging of their components, as our findings suggest. Here, we investigated the effects of sixteen widely used tags on condensate formation in various model organisms, in vitro, in cells and by computational modelling. We find that tagging strongly influenced condensation for some proteins, while others remained unaffected. Effects varied, with some tags enhancing and others decreasing condensation, and depended on the protein being tagged. Coarse-grained simulations suggest that the charge of the fluorescent protein tags is a critical factor modulating condensation behavior. Together, our results underscore the importance of rigorous experimental design and interpretation in condensate experiments.

molecular biology↗