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Bohl, K.

Publications and source records attributed to Bohl, K..

5 recordsLinked to original sources

A VHL-1/HIF-1/SQRD1/COL-88 axis links extracellular matrix formation with longevity in Caenorhabditis elegans

The extracellular matrix (ECM) is a pivotal three-dimensional network crucial for tissue organization, cellular communication, and fundamental cellular processes, where collagens are the major chemical entity in amount. ECM deregulation is directly involved with several pathologies, such as tumour growth and invasiveness, atherosclerosis, and diabetic nephropathy. Mutations in the von Hippel-Lindau tumour suppressor (pVHL) cause VHL syndrome, a multi-tumour syndrome commonly associated with clear cell renal carcinoma (ccRCC). Loss of pVHL is associated with the activation of hypoxia-inducible factor (HIF) signaling. Mutation of VHL-1 in the nematode Canorhabditis elegans has been shown to increase lifespan and stress resistance. Interestingly, considering recent findings on the involvement of collagens in the regulation of lifespan, we also observed these animals to show defects in body morphology in a HIF-1 dependent manner. Based on this finding, we established a link between HIF-1 activation upon loss of VHL-1 and ECM defects associated with alterations in collagen expression. An RNAi screen examining genes upregulated in vhl-1 mutant worms revealed the sulfide quinone oxidoreductase sqrd-1 to mediate the change in body morphology. SQRD-1 is essential to the HIF-1 dependent increase in several collagen genes. One of these genes, col-88, partly mediates both the impact of loss of VHL-1 on lifespan extension and body length. The downregulation of the uncharacterised col-88 partially restores lifespan extension and reduces body size of vhl-1/sqrd-1 to vhl-1(ok161) single mutant. This study contributes to the increasing body of evidence linking lifespan extension and the ECM and now implicates this axis in hypoxia-signaling. These findings are of special interest considering the role of ECM integrity in tumour growth and metastasis. Author SummaryThe extracellular matrix and its composing collagens are associated with a wide number of diseases, including cancer. The von Hippel-Lindau tumour suppressor (pVHL) is known to work by regulating the Hypoxia Inducible Factor (HIF) to help the organism to adapt to lack of oxygen. Mutations in pVHL are associated with clear cell renal carcinoma (ccRCC). Interestingly, a small number of studies have shown that pVHL can be directly associated with collagens, a function that is independent of its classical role regulating HIF. However, there is no further knowledge about which role the hypoxia pathway has when it comes to extracellular matrix formation and function, what would be useful since the invasiveness of cancers, such as ccRCC, are directly connected to their matrix/collagen composition. Here we observed that the model organism C. elegans has drastically different collagen composition and body size upon a mutation on its vhl-1 gene. Furthermore, a protein previously only known to be involved in sulfide metabolism, SQRD-1, connects body size and lifespan in this animal model, revealing a surprising link between the hypoxia pathway and sulfur metabolism to control lifespan. Further studies could target sulfur metabolism in ccRCC to modulate collagen production and tumour invasiveness.

genetics↗

JADE family proteins regulate proteasome abundance and activity

The JADE protein family (JADE1/2/3) has been implicated in a broad range of cellular functions, including WNT signaling, cell cycle control, regulated cell death, and transcriptional regulation through histone acetyltransferase complexes. All three paralogs share high sequence similarity and contain two PHD zinc finger domains. JADE1 has additionally been associated with cilia-related proteins and genetic disorders affecting kidney architecture and function. Despite their widespread expression, the molecular roles of JADE proteins remain incompletely understood. Here, we identify JADE proteins as regulators of proteasome abundance and activity. Using kidney cells as a model, we demonstrate that loss of any single JADE protein led to a marked upregulation of almost all components of the 26S proteasome. Regulation occurred at the post-translational level and was not the consequence of increased transcription. Consistent with a role for JADE proteins in regulating overall proteasomal abundance, JADE-deficient cells displayed elevated proteasomal activity, while ectopic expression of JADE1, JADE2, or JADE3 reduced proteasomal function. Co-immunoprecipitation experiments confirmed the interaction between JADE1 and multiple proteasomal subunits, supporting a direct role of JADE proteins in modulating proteasome turnover, stability and abundance. These findings reveal a novel function of JADE proteins in proteostasis and suggest that their previously reported cellular roles may, in part, be mediated through regulation of the ubiquitin-proteasome system.

cell biology↗

A systematic analysis of diet-induced nephroprotection reveals overlapping and conserved changes in cysteine catabolism

Caloric Restriction (CR) extends lifespan and augments cellular stress-resistance from yeast to primates, making CR an attractive strategy for organ protection in the clinic. Translation of CR to patients is complex, due to problems regarding adherence, feasibility and safety concerns in frail patients. Novel tailored dietary regimens, which modulate the dietary composition of macro- and micronutrients rather than reducing calorie intake promise similar protective effects and increased translatability. However, a direct head-to-head comparison to identify the most potent approach for organ protection as well as overlapping metabolic consequences has not been performed. We systematically analyzed six dietary preconditioning protocols - fasting mimicking diet (FMD), ketogenic diet (KD), dietary restriction of branched chained amino acids (BCAA), two dietary regimens restricting sulfur-containing amino acids (SR80/100) and CR - in a rodent model of renal ischemia-reperfusion injury (IRI) to quantify diet-induced resilience in kidneys. Of the administered diets, FMD, SR80/100 and CR efficiently protect from kidney damage after IRI. Interestingly, these approaches show overlapping changes in oxidative and hydrogen sulfide (H2S)-dependent cysteine catabolism as a potential common mechanism of organ protection. Importantly, these metabolic changes can be recapitulated in patients preconditioned by a diet limiting sulfur-containing amino acids indicating conserved diet-induced mechanisms of stress-resistance that may ultimately lead to clinical application.

biochemistry↗

The proteome of small urinary extracellular vesicles after kidney transplantation as an indicator of renal cellular biology and a source for markers predicting outcome

Kidney transplantation is the preferred renal replacement therapy available. Yet, the biological processes during and after kidney transplantation and how they translate into the overall functional graft outcome are insufficiently understood. Recent developments in the field of extracellular vesicle research allow the deeper exploitation of this non-invasive source. We separated small urinary extracellular vesicles (suEVs) throughout the course of living donor kidney transplantation. SuEVs were collected longitudinally from both the donor and the recipient in 22 living donor kidney transplantations. Unbiased proteomic analysis revealed specific temporal patterns of suEV proteins indicative of the cellular processes involved in the allografts response after transplantation with proteins playing a role in complement activation being among the most dynamically regulated components. Using a leave-one-out cross validation model, we identified potential prognostic markers of kidney function at 1 year after transplantation. One of the proteins identified - phosphoenol pyruvate carboxykinase (PCK2) - could be confirmed in an independent validation cohort of another 22 donor-recipient pairs using targeted mass spectrometry. This study sheds the light on early molecular processes during the course of kidney transplantation and shows the future potential of suEVs as a source of biomarkers in this setting. The data set is provided as a unique resource directly accessible through an online tool that allows dynamic interrogation of this first comprising suEV proteome atlas after kidney transplantation. One Sentence SummaryThis study represents the first atlas of the proteomic changes in small urinary extracellular vesicles throughout living donor kidney transplantation identifying PCK2 abundance as a biomarker for renal function 12 months after transplantation

molecular biology↗

The RNA-binding protein AATF coordinates rRNA maturation

AATF is a central regulator of the cellular outcome upon p53 activation, a finding that has primarily been attributed to its function as a transcription factor. Recent data showed that AATF is essential for ribosome biogenesis and plays a role in rRNA maturation. AATF has been implicated to fulfil this role through direct interaction with rRNA and was identified in several RNA-interactome capture experiments. Here, we provide a first comprehensive analysis of the RNA bound by AATF using CLIP-sequencing. Interestingly, this approach shows predominant binding of the 45S pre-ribosomal RNA precursor molecules. Furthermore, AATF binds to mRNAs encoding for ribosome biogenesis factors as well as snoRNAs. These findings are complemented by an in-depth analysis of the protein interactome of AATF containing a large set of proteins known to play a role in rRNA maturation with an emphasis on the protein-RNA-complexes known to be required for the generation of the small ribosomal subunit (SSU). In line with this finding, the binding sites of AATF within the 45S rRNA precursor localize in close proximity to the SSU cleavage sites. Consequently, our multilayer analysis of the protein-RNA interactome of AATF reveals this protein to be an important hub for protein and RNA interactions involved in ribosome biogenesis.

molecular biology↗