Biology subjects

Harnos, J.

Publications and source records attributed to Harnos, J..

3 recordsLinked to original sources

bioRxiv2025

Armed with PRICKLE(3)s: Stabilizing WNT/PCP complexes against RNF43-mediated ubiquitination

The human Prickle protein family, consisting of PRICKLE1, PRICKLE2, PRICKLE3, and PRICKLE4, is an integral component of the WNT/planar cell polarity (WNT/PCP) pathway and is essential for various cellular and developmental processes. Despite their significance, the detailed roles and involvement in molecular mechanisms of these proteins in cells remain not fully understood. In this study, we used enhanced proximity biotinylation (miniTurboID) combined with mass spectrometry to characterize the microenvironment of PRICKLE1-3. Our results reveal that PRICKLE3 is directly linked to the WNT/PCP pathway, primarily localizing at the plasma membrane and forming complexes with VANGL proteins. This observation prompted us to examine its role in the non-canonical WNT signalling pathway in more detail. Using an inducible expression system to achieve protein levels closer to physiological conditions, we found that PRICKLE3 enhances the stability of VANGL1 and VANGL2 by shielding them from Casein kinase 1 {varepsilon}-mediated phosphorylation. Furthermore, our results indicate that PRICKLE3 modulates WNT receptor complexes by negatively regulating the interaction between Casein kinase 1 {varepsilon} and ubiquitin ligase RNF43, resulting in decreased ubiquitination and increased stabilization of VANGL1/2 at the plasma membrane. Notably, these effects were specific to PRICKLE3, with PRICKLE1 showing no comparable activity. Contrary to previous findings based mainly on standard overexpression studies, neither PRICKLE3 nor PRICKLE1 influenced the levels or phosphorylation status of WNT proteins DISHEVELLED2 and DISHEVELLED3, which are the PRICKLE proteins binding partners. In summary, we have identified a key mechanism specific to PRICKLE3 that positively regulates WNT/PCP complexes by suppressing RNF43. Additionally, we present a comprehensive interactome and new tools for the functional specification of Prickle isoforms to support further research.

cell biology↗

bioRxiv2024

Unveiling Vertebrate Development Dynamics in Frog Xenopus laevis using Micro-CT Imaging

BackgroundXenopus laevis, the African clawed frog, is a versatile vertebrate model organism employed across various biological disciplines, prominently in developmental biology to elucidate the intricate processes underpinning body plan reorganization during metamorphosis. Despite its widespread utility, a notable gap exists in the availability of comprehensive datasets encompassing Xenopus late developmental stages. FindingsIn the present study, we harnessed micro-computed tomography (micro-CT), a non-invasive 3D imaging technique utilizing X-rays to examine structures at a micrometer scale, to investigate the developmental dynamics and morphological changes of this crucial vertebrate model. Our approach involved generating high-resolution images and computed 3D models of developing Xenopus specimens, spanning from premetamorphosis tadpoles to fully mature adult frogs. This extensive dataset enhances our understanding of vertebrate development and is adaptable for various analyses. For instance, we conducted a thorough examination, analyzing body size, shape, and morphological features, with a specific emphasis on skeletogenesis, teeth, and organs like the brain at different stages. Our analysis yielded valuable insights into the morphological changes and structure dynamics in 3D space during Xenopus development, some of which were not previously documented in such meticulous detail. This implies that our datasets effectively capture and thoroughly examine Xenopus specimens. Thus, these datasets hold the solid potential for additional morphological and morphometric analyses, including individual segmentation of both hard and soft tissue elements within Xenopus. ConclusionsOur repository of micro-CT scans represents a significant resource that can enhance our understanding of Xenopus development and the associated morphological changes. The widespread utility of this amphibian species, coupled with the exceptional quality of our scans, which encompass a comprehensive series of developmental stages, opens up extensive opportunities for their broader research application. Moreover, these scans have the potential for use in virtual reality, 3D printing, and educational contexts, further expanding their value and impact. Graphical abstract & lay summary3D images of selected developmental stages of X. laevis in a comparison (scale bar = 10 mm). O_FIG O_LINKSMALLFIG WIDTH=200 HEIGHT=67 SRC="FIGDIR/small/598452v1_ufig1.gif" ALT="Figure 1"> View larger version (14K): org.highwire.dtl.DTLVardef@1bc4664org.highwire.dtl.DTLVardef@1b1750dorg.highwire.dtl.DTLVardef@17bddaaorg.highwire.dtl.DTLVardef@115de4e_HPS_FORMAT_FIGEXP M_FIG C_FIG Lay summaryX-ray tomography was used to examine the African clawed frog (Xenopus laevis). An extensive data set of specimens from tadpoles to adult frogs provides novel insights into the changes and developmental dynamics of selected structures, which opens avenues to an improved understanding of this crucial animal model.

developmental biology↗

bioRxiv2023

A new mechanism of posttranslational polyglutamylation regulates phase separation and signaling of the Wnt pathway protein Dishevelled.

Polyglutamylation is a reversible post-translational modification that is catalyzed by enzymes from the tubulin tyrosine ligase-like (TTLL) family. Here, we found that TTLL11 generates a previously unknown type of polyglutamylation initiated by the addition of a glutamate residue to the free C-terminal carboxyl group of a substrate protein. TTLL11 efficiently polyglutamylates the Wnt signaling protein Disheveled 3 (DVL3), thereby changing the interactome of DVL3, as well as it increases its capacity to get phosphorylated, to undergo liquid-liquid phase separation (LLPS), and to act in the non-canonical Wnt pathway. Both carboxyterminal polyglutamylation and the resulting reduction in LLPS capacity of DVL3 were reverted by the deglutamylating enzyme CCP6, which demonstrates the causal relationship between TTLL11-mediated polyglutamylation and LLPS. We thus discovered a novel type of posttranslational modification, which significantly broadens the range of proteins that can be modified by polyglutamylation and provides the first evidence that polyglutamylation can act as a regulator of protein LLPS.

cell biology↗