Biology subjects

Voloshanenko, O.

Publications and source records attributed to Voloshanenko, O..

4 recordsLinked to original sources

bioRxiv2025

Secretion and signaling properties of Wnt11 and Wnt3 are determined by their N-terminus

Wnt proteins are evolutionarily conserved signaling molecules that control cell-cell communication in health and disease. While different Wnt proteins share a requirement for palmitoleoylation and the cargo receptor Evi/Wls for their secretion, they interact with diverse signaling receptors and co-receptors to elicit distinct outcomes in signal-receiving cells. The molecular mechanisms underlying this contrast between conserved post-translational modification and secretion and diverse receptor interactions remain poorly understood. Here, we demonstrate that Wnt11 maintains partial secretion capability in the absence of Evi/Wls. Unexpectedly, the secreted protein is also non-palmitoleoylated, yet retains its ability to activate downstream signaling in receiving cells through the Frizzled 6 receptor. We identify the N-terminus as the mediator of this unusual behavior, as Wnt3, which strictly requires Evi/Wls, becomes Evi/Wls-independent when its N-terminus is replaced with that of Wnt11. In different cell lines and in Xenopus laevis embryos, the chimeric Wnt11Nterm-Wnt3 protein recapitulates known Wnt11 phenotypes, demonstrating that the N-terminal portion of Wnt11 is sufficient to determine signaling specificity. Our findings reveal novel determinants of Wnt protein specificity and provide mechanistic insights into the molecular basis of Wnt secretion and downstream signaling.

cell biology↗

bioRxiv2025

Wnt10b signaling regulates replication stress-induced chromosomal instability in human cancer

Wnt signaling pathways are involved in various developmental and tissue maintenance functions while deregulated Wnt signaling is closely linked to human cancer. Recent work revealed that loss of Wnt signaling impairs mitosis and causes abnormal microtubule growth at the mitotic spindle resulting in chromosome missegregation and aneuploidy, both of which are hallmarks of cancer cells exhibiting chromosomal instability (CIN). Here, we demonstrate that Wnt signaling specifically activated by Wnt10b is relevant in colorectal cancer cells to prevent abnormal microtubule dynamics and chromosome missegregation. Although mitosis is affected, Wnt10b signaling is required during the S phase of the cell cycle. In fact, Wnt10b signaling acts upon DNA replication stress in S phase, a condition typically associated with CIN in cancer, to prevent increased microtubule dynamics from S phase until mitosis, where they otherwise manifest in chromosome missegregation. Interestingly, replication stress-induced chromosomal breaks are also efficiently suppressed by Wnt10b. Thus, our results show that Wnt10b signaling regulates replication stress-induced chromosome missegregation and breakage, and hence, is a determinant for broad genome instability in cancer cells. Summary blurbWe describe a novel role of Wnt10b signaling acting in response to DNA replication stress to suppress chromosomal breaks and mitotic errors in human cancer cells.

cell biology↗

bioRxiv2025

Image-based screens identify regulators of endogenous Dvl2 biomolecular condensates

Dishevelled (Dvl) proteins are essential transducers in Wnt signaling pathways, which have been implicated in development, stem cell maintenance, and human diseases such as cancer. Several studies have shown that Dvl proteins form dynamic biomolecular condensates. However, how cellular signals and cell states influence the formation of biomolecular condensates remains poorly understood. Here, we analyzed cells with endogenous Dvl2 condensates using image-based cell sorting in combination with phosphoproteomics and identified protein enrichment for Wnt/PCP signaling and the G2/M cell cycle transition. We then performed an image-based high-throughput screen to identify small molecule kinase inhibitors that affect Dvl2 liquid-liquid phase separation. Strikingly, CK1{delta}/{varepsilon} inhibition blocked Dvl2 condensate formation. Its effect on Wnt signaling was modulated in genetic epistasis experiments with loss-of-function alleles of APC, Axin1, and MCC. Our study highlights the interplay between post-translational modifications and condensate dynamics, opening new avenues for research on their role in cellular signaling and disease intervention.

cell biology↗

bioRxiv2020

Allele-specific endogenous tagging and quantitative analysis of β-catenin in colon cancer cells

Wnt signaling plays important roles in development, homeostasis, and tumorigenesis. Mutations in {beta}-catenin that activate Wnt signaling have been found in colorectal and hepatocellular carcinomas. However, the dynamics of wild-type and mutant forms of {beta}-catenin are not fully understood. Here, we genome-engineered fluorescently tagged alleles of the endogenous {beta}-catenin in a colorectal cancer cell line. Wild-type and oncogenic mutant alleles were tagged with different fluorescent proteins, enabling the analysis of both variants in the same cell. We analyzed the properties of both {beta}-catenin alleles using immunoprecipitation, immunofluorescence and fluorescence correlation spectroscopy approaches, revealing distinctly different biophysical properties. In addition, activation of Wnt signaling by treatment with a GSK3{beta} inhibitor or a truncating APC mutation modulated the wild-type allele to mimic the properties of the mutant {beta}-catenin allele. The one-step tagging strategy demonstrates how genome engineering can be employed for the parallel functional analysis of different genetic variants.

genetics↗