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Preisinger, C.

Publications and source records attributed to Preisinger, C..

3 recordsLinked to original sources

A microbiota-derived protease links phage susceptibility to host epithelial responses

Bacteriophages are major ecological drivers of gut microbial ecology, yet whether bacterial mechanisms that determine phage susceptibility have consequences for the mammalian host remains poorly understood. Here, we identify dipeptidyl peptidase 11 (Dpp11a), the predominant active serine protease of the prevalent gut commensal Phocaeicola vulgatus, as an unexpected bacterial defence factor. Dpp11a protects against environmental proteases and confers resistance to bacteriophage infection. Metatranscriptomic analyses further reveal increased expression of both dpp11a and P. vulgatus-associated phage transcripts in ulcerative colitis stool samples, indicating that both components of this interaction are transcriptionally active in disease-associated human microbiomes. Using the microfluidic gut-on-a-chip co-culture model HuMiX, we show that the absence of Dpp11 is accompanied by altered epithelial tight-junction remodelling during phage-bacterial infection. Together, our findings reveal that the consequences of bacterial phage defence can extend beyond phage-bacterium interactions to the mammalian epithelium.

microbiology↗

GAPVD1 regulates CSNK1D-dependent PER2 FASP phosphorylation

The mammalian circadian clock relies on precisely timed transcriptional and post-translational events to generate [~]24-hour rhythms. A key post-translational mechanism is phosphorylation of PER2 by CSNK1D, governed by a phosphoswitch that integrates stabilizing and destabilizing phosphorylation marks at the FASP and Degron sites to control PER2 stability and circadian period. Here, a novel role for the PER complex protein GAPVD1 in regulating CSNK1D-mediated FASP site phosphorylation is revealed. GAPVD1 associates with PER2 and CSNK1D via a distinct helical bundle domain, weakening PER2-CSNK1D association and attenuating PER2 FASP phosphorylation in cells and in vitro. Mechanistically, this helical bundle interacts with the PER2 casein kinase binding domain and is contacted by the adjacent VPS9 region in GAPVD1, providing a structural explanation for how flanking GAPVD1 domains can control access of binding proteins to the helical bundle. Notably, a minimal GAPVD1 helical bundle fragment is sufficient for binding to PER2 and CSNK1D but not for robust attenuation of PER2 FASP phosphorylation, indicating that additional regions of GAPVD1 are required. GAPVD1-mediated inhibition of PER2 FASP phosphorylation also turned out to be independent of the CSNK1D C-terminal tail, pointing to mechanisms such as displacement of PER2 FASP from the CSNK1D active site or allosteric regulation of CSNK1D substrate engagement. Together, these findings identify GAPVD1 as a PER-complex scaffold that modulates the PER2-CSNK1D control node and introduce an additional regulatory layer into the circadian phosphoswitch. A GAPVD1 helical bundle domain binds the PER2 CKBD and CSNK1D to weaken PER2-CSNK1D interaction and attenuate PER2 FASP phosphorylation.

molecular biology↗

The nucleoporin Nup50 activates the Ran guanyl-nucleotide exchange factor RCC1 to promote mitotic NPC assembly

During mitotic exit, thousands of nuclear pore complexes (NPCs) assemble concomitant with the nuclear envelope to build a transport-competent nucleus. We show here that Nup50 plays a crucial role in NPC assembly that is independent of its well-established function in nuclear transport. RNAi-mediated downregulation in cells or immunodepletion of the protein in Xenopus egg extracts interferes with NPC assembly. We define a conserved central region of 46 residues in Nup50 that is crucial for Nup153 and MEL28/ELYS binding, and NPC interaction. Surprisingly, neither NPC interaction nor binding of Nup50 to importin , {beta}, the GTPase Ran or chromatin is crucial for its function in the assembly process. Instead, we discovered that an N-terminal fragment of Nup50 can stimulate the Ran guanine exchange factor RCC1 and NPC assembly, indicating that Nup50 acts via the Ran system in mitotic NPC reformation. In support of this conclusion, Nup50 mutants defective in RCC1 binding and stimulation cannot replace the wild type protein in in vitro NPC assembly assays.

cell biology↗