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Rämisch, S.

Publications and source records attributed to Rämisch, S..

3 recordsLinked to original sources

Targeting the translation machinery: The RNase Y specificity factor Y-complex coordinates ribosome degradation

Limiting the synthesis and activity of ribosomes is crucial for adaptation to stresses, such as heat or nutrient starvation. In Bacillus subtilis, this can be achieved through the coordinated action of the alarmones (p)ppGpp and the transcription factor Spx. Here, we performed a genetic screen to uncover novel factors contributing to the heat shock response of B. subtilis. We identified the Y-complex, which confers specificity to the endonuclease RNase Y, as a critical player during stress conditions, such as heat or transition into stationary phase. This protein complex is required for processing diverse RNAs, notably the maturation of mRNAs encoding proteins involved in translation and metabolism. We further demonstrate that the Y-complex and RNase Y initiate the degradation of rRNAs of mature ribosomes, lowering their abundance. We propose that the Y-complex is a regulatory hub that modulates gene expression, adjusts protein synthesis and resource allocation.

microbiology↗

Protein arginine phosphorylation and de-phosphorylation facilitate protein homeostasis by an AAA+ chaperone protease system

We could demonstrate that the AAA+ unfoldase ClpC together with the protein arginine kinase and adaptor protein McsB, its activator McsA and the phosphatase YwlE form a unique chaperone system. Here, the McsA-activated McsB phosphorylates and targets aggregated substrate proteins for extraction and unfolding by ClpC. Sub-stoichiometric amounts of the YwlE phosphatase enhanced the ClpC/McsB/McsA mediated disaggregation and facilitated the de-phosphorylation of the arginine-phosphorylated substrate protein extruded by ClpC, allowing its subsequent refolding. Interestingly, the successfully refolded protein escaped degradation by the loosely associated ClpP protease. This unique chaperone system is thereby able to disaggregate and refold aggregated proteins but can also remove severely damaged protein aggregates by degradation.

microbiology↗

DnaK response to expression of protein mutants is dependent on translation rate and stability

Chaperones play a central part in the quality control system in cells by clearing misfolded and aggregated proteins. The chaperone DnaK acts as a sensor for molecular stress by recognising short hydrophobic stretches of misfolded proteins. As the level of unfolded protein is a function of protein stability, we hypothesised that the level of DnaK response upon overexpression of recombinant proteins would be correlated to stability. Using a set of mutants of the{lambda} -repressor with varying thermal stabilities and a fluorescent reporter system, the effect of stability on DnaK response and protein abundance was investigated. Our results demonstrate that the initial DnaK response is largely dependent on protein synthesis rate but as the recombinantly expressed protein accumulates and homeostasis is approached the response correlates strongly with stability. Furthermore, we observe a large degree of cell-cell variation in protein abundance and DnaK response in more stable proteins.

biochemistry↗