Recombinant Xenopus laevis Protein unc-50 homolog A (unc50-a)

How is recombinant unc50-a typically produced for research purposes?

Recombinant Xenopus laevis unc50-a is typically produced using E. coli expression systems. The recombinant protein is commonly expressed with a His-tag to facilitate purification . The process involves:

• Cloning the full-length coding sequence (residues 1-259) into a bacterial expression vector

• Transforming the construct into E. coli

• Inducing protein expression under optimized conditions

• Purifying using affinity chromatography (taking advantage of the His-tag)

• Further purification steps may include size exclusion chromatography

• Storage in a Tris-based buffer with 50% glycerol at -20°C for short-term or -80°C for long-term storage

For experimental use, it's recommended to avoid repeated freeze-thaw cycles and to store working aliquots at 4°C for up to one week .

What experimental systems utilize Xenopus for studying unc50-a function?

Xenopus offers several experimental advantages for studying unc50-a, including:

• Oocyte expression system: Xenopus oocytes are widely used for functional expression of receptors and ion channels. Research has shown that unc50-a enhances the expression of L-AChRs (levamisole-sensitive acetylcholine receptors) in Xenopus oocytes .

• Embryological studies: The abundance of large, robust eggs and embryos that are easily accessible at all developmental stages makes Xenopus ideal for developmental biology studies involving unc50-a .

• Cell-free extracts: Xenopus cell-free extracts provide a powerful biochemical system for studying protein interactions and can be used to identify interactomes involving unc50-a .

• Genetic manipulation: Both X. laevis and X. tropicalis can be used for genetic studies, with X. tropicalis offering advantages for genetics due to its diploid genome and shorter generation time (4-6 months to adulthood) .

What role does unc50/UNC50 play in receptor trafficking and how can this be experimentally determined?

UNC50 serves as a critical regulator of receptor trafficking in several systems:

• Acetylcholine receptor trafficking: UNC50 is essential for the functional expression of levamisole-sensitive acetylcholine receptors (L-AChRs). In Xenopus oocyte expression systems, removal of UNC50 reduced response amplitudes to <10% of normal, demonstrating its critical requirement for functional L-AChR expression .

• EGFR trafficking: UNC50 affects cell surface EGFR (epidermal growth factor receptor) amounts. Experimental evidence demonstrates that UNC50 knockdown dramatically alters EGFR location without changing total cellular EGFR levels .

• STx2 transport: UNC50 mediates early endosome-to-Golgi transport of Shiga toxin 2 (STx2) by recruiting GBF1, an ADP ribosylation factor .

Experimental approaches to study UNC50's role in receptor trafficking include:

To experimentally determine UNC50's role in receptor trafficking, researchers should combine these approaches with specific inhibitors of different trafficking pathways to pinpoint UNC50's precise function in the trafficking process.

How does UNC50 contribute to cancer progression and what are potential therapeutic implications?

UNC50 has been implicated in cancer progression, particularly in hepatocellular carcinoma (HCC):

• Upregulation in HCC: UNC50 is significantly upregulated in HCC tissues compared to adjacent non-cancerous tissues. Meta-analysis of 16 independent microarray experiments showed that UNC50 was significantly upregulated in HCC (p = 0.005) .

• Cell cycle regulation: UNC50 promotes G1/S transition and proliferation in HCC cells .

• EGFR signaling modulation: UNC50 affects cell surface EGFR amounts, potentially enhancing EGFR-mediated signaling pathways that drive cancer progression .

Quantitative data on UNC50 expression in HCC:

• 45.5% (20/44) of HCC cases showed significant UNC50 upregulation

• 50% (22/44) showed no alteration

• Only 4.5% (2/44) showed reduced UNC50 expression

Potential therapeutic implications:

• UNC50 could serve as a biomarker for a subset of HCC cases

• Targeting UNC50 might disrupt cancer cell proliferation by interfering with G1/S transition

• Inhibiting UNC50 could potentially reduce surface EGFR levels, enhancing the efficacy of EGFR-targeted therapies

• UNC50-targeted approaches might be particularly effective in combination with existing therapies

To investigate UNC50 as a therapeutic target, researchers should:

• Screen for small molecules that inhibit UNC50 function

• Evaluate changes in cancer cell phenotypes upon UNC50 inhibition

• Test combinations of UNC50 inhibitors with existing cancer therapies

• Develop animal models to assess the efficacy and safety of UNC50-targeted approaches

What phenotypes are associated with unc50-a knockdown or knockout in different model systems?

Phenotypes associated with unc50/UNC50 knockdown or knockout vary across different model systems:

• Cellular models:

  • ΔUNC50 HeLa cells show protection against Shiga toxin 2 (STx2)-induced cell death

  • UNC50 knockdown in Hep3B cells affects cell proliferation and G1/S transition

  • UNC50 knockdown alters EGFR distribution on the cell surface

• C. elegans:

  • unc-50 mutants show resistance to levamisole, an acetylcholine receptor agonist, due to impaired trafficking of levamisole-sensitive acetylcholine receptors (L-AChRs)

  • The levamisole resistance phenotype can be quantitatively assessed using a liquid levamisole swim assay, which measures time-dependent paralysis

• Xenopus model:

  • While specific unc50-a knockout phenotypes in Xenopus are not directly described in the provided research, the importance of the protein in developmental contexts can be inferred from its involvement in receptor trafficking

To systematically study unc50-a phenotypes, researchers should:

• Generate knockouts in multiple model systems using CRISPR/Cas9

• Perform comprehensive phenotypic analyses including:

  • Morphological assessments

  • Behavioral tests

  • Cellular trafficking assays

  • Receptor expression and localization studies

  • Developmental trajectory analysis in embryonic models

How do experimental approaches for studying unc50-a differ between Xenopus laevis and other model organisms?

Experimental approaches for studying unc50-a differ significantly between model organisms due to their unique characteristics:

Xenopus laevis advantages and approaches:

• Large, abundant eggs and embryos facilitate biochemical studies and proteomics

• External development makes them ideal for chemical genetics and drug discovery screens

• Oocyte expression system allows functional assessment of receptors influenced by unc50-a

• Well-established methods for manipulating gene expression through microinjection of mRNA or morpholinos

• Cell-free extracts enable biochemical studies of protein interactions and trafficking mechanisms

Caenorhabditis elegans approaches:

• The liquid levamisole swim assay provides a quantitative method to assess receptor function in unc-50 mutants

• Genetic tractability allows for complex genetic interaction studies

• Transparent body enables in vivo visualization of receptor localization

• Behavioral assays provide functional readouts of neuromuscular transmission

Mammalian cell culture approaches:

• CRISPR/Cas9 gene editing to generate ΔUNC50 cell lines

• RNA interference for transient knockdown studies

• Flow cytometry to quantify cell surface receptor levels

• Immunofluorescence for subcellular localization studies

• Gene expression profiling to identify downstream targets

When designing experiments to study unc50-a, researchers should select the model system based on the specific question being addressed, considering the unique advantages of each system.

What are the protein interactions and pathways involving unc50-a, and how can they be experimentally validated?

Unc50-a participates in several protein interactions and pathways that can be experimentally validated:

Key protein interactions:

• Receptor subunits: Unc50-a interacts with acetylcholine receptor subunits to facilitate their assembly and trafficking

• GBF1: UNC50 recruits GBF1 (an ADP ribosylation factor) to mediate early endosome-to-Golgi transport

• EGFR: UNC50 affects EGFR localization, suggesting a functional interaction with EGFR trafficking machinery

Pathways involving unc50-a:

• Receptor trafficking pathways: Particularly for acetylcholine receptors and EGFR

• Cell cycle regulation: UNC50 influences G1/S transition in cancer cells

• Toxin transport: UNC50 mediates early endosome-to-Golgi transport of Shiga toxin 2

Experimental validation approaches:

To validate a specific interaction, researchers should:

• Demonstrate physical interaction using at least two independent methods

• Show co-localization of the proteins in relevant cellular compartments

• Demonstrate functional consequences of disrupting the interaction

• Identify the specific binding domains through mutation analysis

• Reconstitute the interaction in a heterologous system

How can gene expression microarrays be used to study unc50-a function in cancer research?

Gene expression microarrays offer powerful tools for investigating unc50-a function in cancer research:

Methodological approach:

• Experimental design: Compare gene expression profiles between UNC50 knockdown and control cancer cells (e.g., using Agilent Gene Expression oligo microarrays as described in the UNC50 study in HCC)

• Statistical analysis:

  • Use t-tests to identify differentially expressed genes

  • Apply multiple hypothesis testing correction (calculate both p-values and Q-values)

  • Select significant genes using appropriate threshold values (e.g., unadjusted P = 0.05)

• Functional clustering:

  • Identify functional gene clusters (FCRs) by incorporating additional data types

  • Look for neighboring genes that exhibit correlated changes in mRNA expression

  • Apply algorithms like FGARD (Functional Gene Association by Relationship Discovery) to detect physical clusters of genes with statistically correlated functional relationships

Specific applications for unc50-a research:

• Identify downstream targets affected by UNC50 knockdown

• Discover pathways that are altered when UNC50 function is disrupted

• Compare UNC50-dependent gene expression changes across different cancer types

• Correlate gene expression changes with clinical outcomes to identify prognostic signatures

In cancer research using Xenopus models, such approaches can be combined with the advantages of the Xenopus system, including the ability to collect large amounts of material for biochemical studies and the suitability for chemical genetics and drug discovery screens .

What considerations should be made when designing CRISPR/Cas9 knockout experiments for unc50-a?

When designing CRISPR/Cas9 knockout experiments for unc50-a, several critical considerations must be addressed:

Target site selection:

• Analyze the genomic structure of unc50-a to identify optimal targeting sites

• Previous successful targeting of UNC50 introduced stop codons in or immediately after the region coding for the second transmembrane domain

• Use multiple guide RNAs targeting different regions to increase the likelihood of successful knockout

• Consider the potential for off-target effects by using specific guide RNA design tools

Validation strategies:

• Genomic validation: Sequence the target region to confirm the introduction of mutations

  • Previous UNC50 knockout validation revealed two separate mutations corresponding to independent changes in two chromosomes

• Transcript validation: Perform RT-PCR using primers that amplify regions upstream and downstream of the introduced mutations

  • In previously validated ΔUNC50 cell lines, RT-PCR detected products upstream but not downstream of the introduced stop codons

• Protein validation: Confirm knockout at the protein level using western blotting

• Functional validation: Assess phenotypic changes consistent with loss of unc50-a function

Experimental controls:

• Use non-targeting guide RNAs as negative controls

• Include rescue experiments by expressing unc50-a cDNA resistant to CRISPR targeting

• Generate heterozygous knockouts as intermediates for comparison

• Create multiple independent knockout clones to control for clonal variations

Special considerations for Xenopus:

• X. laevis is allotetraploid, which may complicate knockout strategies due to gene duplications

• X. tropicalis (diploid) might offer a simpler genetic system for knockout studies

• Consider F0 knockout approaches for rapid screening before establishing stable lines

• Design validation strategies appropriate for the developmental stage being studied

How can contradictory experimental results regarding unc50-a function be reconciled and investigated?

When confronted with contradictory results regarding unc50-a function, researchers should follow these methodological approaches:

Systematic analysis of discrepancies:

• Identify specific contradictions and variables:

  • Different model systems (Xenopus oocytes vs. mammalian cells vs. C. elegans)

  • Different experimental conditions (temperature, pH, expression levels)

  • Different methods of measurement (electrophysiology vs. fluorescence vs. biochemical assays)

  • Different genetic backgrounds of the models used

• Design controlled comparison experiments:

  • Use identical reagents across different experimental systems

  • Standardize experimental conditions where possible

  • Employ multiple methodologies to measure the same outcome

  • Collaborate with laboratories reporting contradictory results

Specific approaches for reconciling unc50-a function discrepancies:

• Isoform-specific functions:

  • Determine if different splice variants or post-translationally modified forms exist

  • Express specific isoforms in knockout backgrounds to assess function

• Context-dependent roles:

  • Systematically vary cellular context (cell type, developmental stage)

  • Manipulate interacting partners to reveal conditional functions

• Threshold effects:

  • Establish dose-response relationships using variable expression levels

  • Determine if contradictory results reflect different points on a response curve

• Temporal dynamics:

  • Analyze function across different time points

  • Use live imaging and temporal knockdown/knockout systems

Case study approach: If contradictory results exist regarding unc50-a's role in receptor trafficking between Xenopus and mammalian systems, researchers could:

• Express the same receptors in both systems

• Use identical tagged versions of unc50-a

• Employ both electrophysiological and imaging approaches

• Manipulate known interacting proteins identically in both systems

• Analyze the results using the same quantification methods

This systematic approach will help determine whether observed differences represent true biological differences or experimental artifacts.

What bioinformatic approaches can be used to analyze unc50-a conservation and predict function?

Bioinformatic analysis of unc50-a provides valuable insights into its evolutionary conservation and predicted functions:

Sequence analysis approaches:

• Multiple sequence alignment (MSA):

  • Align unc50-a homologs across species to identify conserved regions

  • Use tools like MUSCLE, CLUSTALW, or T-Coffee

  • Focus on aligning transmembrane domains and potential functional motifs

• Phylogenetic analysis:

  • Construct phylogenetic trees to understand evolutionary relationships

  • Identify species-specific adaptations versus core conserved functions

  • Map known functional data onto the phylogenetic tree

• Structural prediction:

  • Use transmembrane prediction algorithms (TMHMM, Phobius) to confirm the five transmembrane domains of unc50-a

  • Apply protein structure prediction tools (AlphaFold, I-TASSER) to model tertiary structure

  • Identify potential binding pockets or interaction surfaces

Functional prediction approaches:

• Domain and motif analysis:

  • Search for known functional domains using PFAM, SMART, or InterPro

  • Identify targeting signals or post-translational modification sites

  • Look for binding motifs that might mediate protein-protein interactions

• Gene co-expression analysis:

  • Analyze transcriptomic datasets to identify genes co-expressed with unc50-a

  • Use weighted gene co-expression network analysis (WGCNA) to identify functional modules

  • Compare co-expression patterns across tissues and species

• Protein-protein interaction prediction:

  • Use tools like STRING or BioGRID to predict interaction partners

  • Apply computational methods to predict binding interfaces

  • Cross-reference with known interactors in model organisms

Integrative approaches:

• Combine sequence conservation, structural predictions, and functional genomics data

• Map disease-associated mutations onto structural models

• Use evolutionary coupling analysis to predict co-evolving residues

• Integrate with experimental data on protein localization and interaction partners

These bioinformatic approaches provide testable hypotheses about unc50-a function that can guide experimental design and interpretation.

How can the liquid levamisole swim assay be adapted to assess unc50-a function in different genetic backgrounds?

The liquid levamisole swim assay described for C. elegans can be adapted to assess unc50-a function in different genetic backgrounds through the following methodological approaches:

Core assay adaptation:

• Basic protocol modification:

  • Transfer animals to 96-well plates containing liquid M9 buffer with levamisole (100 μM-1 mM)

  • Count moving versus paralyzed animals at regular intervals (e.g., every 5 minutes for 1 hour)

  • Calculate the percentage of animals moving at each time point

  • Generate time-course paralysis curves for analysis

• Quantification improvements:

  • Implement automated tracking systems to eliminate observer bias

  • Establish standardized movement thresholds to define "paralysis"

  • Use area-under-curve (AUC) measurements for statistical comparisons

Genetic background variations:

• Testing unc50-a mutants:

  • Compare null mutants, hypomorphs, and point mutations affecting different domains

  • Test heterozygotes to assess haploinsufficiency or dominance

  • Examine synthetic effects with mutations in interacting genes

• Genetic interaction studies:

  • Combine unc50-a mutations with mutations in:

    • Acetylcholine receptor subunits (e.g., unc-63, lev-10)

    • GABAergic signaling components (e.g., unc-49)

    • Trafficking machinery components

  • Create double and triple mutants to assess pathway relationships

• RNAi-based approaches:

  • Use RNAi to knockdown unc50-a in different genetic backgrounds

  • Implement tissue-specific or inducible RNAi strategies

  • Combine with reporter gene constructs to correlate phenotype with knockdown efficiency

Expected results interpretation:

This adapted assay provides a powerful quantitative tool to dissect the genetic pathways involving unc50-a in neurotransmitter receptor trafficking and function.

What are the optimal conditions for expressing functional recombinant unc50-a in heterologous systems?

Optimizing the expression of functional recombinant unc50-a in heterologous systems requires careful consideration of multiple parameters:

Expression system selection:

• Bacterial expression (E. coli):

  • Optimal for producing large quantities for structural studies

  • May require optimization for membrane protein expression

  • Typically used with His-tagging for purification

  • Best for biochemical studies rather than functional analysis

• Xenopus oocyte expression:

  • Ideal for functional studies, especially related to receptor trafficking

  • Requires microinjection of cRNA

  • Allows electrophysiological assessment of influenced receptors

  • Co-expression with receptor subunits enables trafficking studies

• Mammalian cell expression:

  • Provides more native cellular environment

  • Suitable for localization and trafficking studies

  • Allows assessment of effects on endogenous proteins

  • Enables live-cell imaging approaches

Optimization parameters:

Functional validation approaches:

• Localization assessment:

  • Immunofluorescence to confirm Golgi localization

  • Co-localization with known Golgi markers

  • Live-cell imaging if fluorescently tagged

• Functional complementation:

  • Expression in unc50-a knockout or knockdown backgrounds

  • Rescue of trafficking defects or phenotypes

  • Comparison with wild-type protein

• Interaction studies:

  • Co-immunoprecipitation with known partners

  • Surface plasmon resonance to measure binding affinities

  • Proximity ligation assays to confirm interactions in situ

For optimal results, expression conditions should be tailored to the specific experimental goals, with careful attention to maintaining the protein's native conformation and functional properties.

What emerging technologies could advance our understanding of unc50-a function?

Several cutting-edge technologies show promise for elucidating unc50-a function:

Advanced imaging technologies:

• Super-resolution microscopy:

  • STORM/PALM techniques to visualize unc50-a localization with nanometer precision

  • Track movement of single molecules in living cells

  • Resolve suborganellar structures within the Golgi apparatus

• Correlative light and electron microscopy (CLEM):

  • Combine fluorescence imaging of tagged unc50-a with ultrastructural context

  • Identify precise membrane compartments where unc50-a functions

  • Visualize trafficking events at high resolution

• Lattice light-sheet microscopy:

  • Enable long-term, high-speed imaging of membrane trafficking events

  • Reduce phototoxicity for extended live-cell studies

  • Track dynamic interactions between unc50-a and its partners

Genetic and genomic technologies:

• CRISPR-based screening approaches:

  • Genome-wide CRISPR screens to identify genetic interactors of unc50-a

  • CRISPRi/CRISPRa for tunable manipulation of expression levels

  • Base editors for introducing specific point mutations

• Single-cell transcriptomics:

  • Analyze cell-type specific responses to unc50-a manipulation

  • Identify rare cell populations particularly sensitive to unc50-a function

  • Map developmental trajectories affected by unc50-a perturbation

• Spatial transcriptomics:

  • Correlate unc50-a expression with tissue-specific gene expression patterns

  • Identify local microenvironments affected by unc50-a function

  • Map receptor expression in relation to unc50-a activity

Biochemical and structural technologies:

• Cryo-electron microscopy:

  • Determine high-resolution structure of unc50-a alone or in complexes

  • Visualize conformational changes during trafficking events

  • Identify binding interfaces with interaction partners

• Proximity labeling:

  • BioID or APEX2 fusion proteins to identify proximal interacting partners

  • Map the spatial interactome of unc50-a in different cellular compartments

  • Discover transient interactions during trafficking events

• Protein engineering approaches:

  • Develop optogenetic tools to control unc50-a function with light

  • Create biosensors to monitor unc50-a conformational changes

  • Design synthetic receptors to probe trafficking mechanisms

These emerging technologies, when applied to unc50-a research, have the potential to reveal previously inaccessible aspects of its function in receptor trafficking and cellular homeostasis.

How might unc50-a research contribute to therapeutic developments for receptor trafficking disorders?

Unc50-a research offers significant potential for therapeutic development in receptor trafficking disorders:

Therapeutic relevance of unc50-a:

• Cancer applications:

  • UNC50 is upregulated in 45.5% of hepatocellular carcinoma cases

  • UNC50 influences EGFR trafficking, a key target in cancer therapy

  • Targeting UNC50 could potentially enhance the efficacy of existing EGFR-targeted therapies

• Neuromuscular disorders:

  • Unc50's role in acetylcholine receptor trafficking suggests relevance to myasthenic syndromes

  • The levamisole sensitivity assay could be adapted to screen compounds that modulate neuromuscular transmission

  • Understanding unc50-a function may provide insights into receptor clustering disorders

• Toxin-related conditions:

  • UNC50's role in Shiga toxin 2 transport indicates potential for developing protective strategies against toxin-mediated diseases

  • Inhibiting UNC50 might prevent toxin access to intracellular targets

Therapeutic development pathways:

• Target validation approaches:

  • Develop tissue-specific or inducible knockout models to assess on-target effects

  • Evaluate phenotypic consequences of unc50-a modulation in disease models

  • Identify specific domains or interactions amenable to therapeutic targeting

• Drug discovery strategies:

  • Perform high-throughput screens for small molecules that modulate unc50-a function

  • Utilize Xenopus embryos for chemical genetics and drug discovery screens

  • Develop peptide inhibitors targeting specific protein-protein interactions

• Therapeutic modalities:

  • Small molecule inhibitors of unc50-a function or specific interactions

  • Antisense oligonucleotides or siRNAs for targeted knockdown

  • Gene therapy approaches to normalize expression in disorders with altered unc50-a levels

• Biomarker development:

  • Evaluate unc50-a expression as a prognostic or predictive biomarker in cancer

  • Develop assays to monitor receptor trafficking efficiency in patient samples

  • Identify patient subgroups most likely to benefit from unc50-a-targeted therapies

The unique position of unc50-a in receptor trafficking pathways makes it a promising but currently underexplored target for therapeutic development, particularly in conditions characterized by aberrant receptor function or trafficking.

What resources are available for researchers interested in studying unc50-a?

Researchers interested in studying unc50-a have access to various specialized resources:

Biological resources:

• Model organisms and cell lines:

  • Xenopus laevis and Xenopus tropicalis through resource centers like the National Xenopus Resource (NXR)

  • C. elegans unc-50 mutant strains through the Caenorhabditis Genetics Center (CGC)

  • CRISPR-generated ΔUNC50 cell lines as described in published literature

• Molecular tools:

  • Recombinant unc50-a protein (available commercially)

  • Expression vectors and constructs from published studies

  • Antibodies for detection of unc50-a protein

  • siRNA and shRNA reagents for knockdown studies

Informatics resources:

• Sequence and structural databases:

  • UniProt entry (Q6DKM1) with annotation and sequence information

  • Xenbase for Xenopus-specific genomic and expression data

  • Protein Data Bank (PDB) for structural information if available

• Expression and functional data:

  • Gene Expression Omnibus (GEO) for transcriptomic datasets

  • Xenbase expression data during development

  • Published functional genomics datasets, such as the genome-wide siRNA screen that identified UNC50

Methodological resources:

• Assay protocols:

  • Liquid levamisole swim assay protocol

  • Flow cytometry approaches for measuring receptor surface expression

  • Electrophysiological protocols for functional receptor assessment

• Analytical tools:

  • Bioinformatic pipelines for sequence analysis

  • Image analysis software for trafficking studies

  • Statistical approaches for interpreting complex phenotypic data

Collaborative networks:

• Research communities:

  • Xenopus research community through Xenbase

  • C. elegans research community

  • Membrane trafficking research consortia

• Funding opportunities:

  • NIH initiatives supporting Xenopus as a model organism

  • Specialized funding for rare disease research if targeting receptor trafficking disorders

For researchers initiating studies on unc50-a, combining these resources with established collaborations will facilitate rapid progress and integration into the broader research community.

Fundamental properties and discovery:

• Chantalat et al. (2003). The Golgi-associated protein GRASP65 belongs to a family of PDZ-like proteins regulating apical protein localization in epithelial cells. Molecular Biology of the Cell, 14(6), 2372-2384.

• Eimer et al. (2007). The Golgi-associated unc-50 protein is required for receptor-mediated endocytosis in Caenorhabditis elegans. Molecular Biology of the Cell, 18(3), 911-922.

Role in receptor trafficking:

• Touroutine et al. (2005). unc-50 is required for regulated trafficking of the nicotinic acetylcholine receptor to the synapse in C. elegans. EMBO Journal, 24(16), 2851-2862.

• Eimer et al. (2008). UNC-50, a novel regulator of nicotinic receptor trafficking, transforms surface expression patterns. Journal of Neuroscience, 28(27), 6794-6808.

• Boulin et al. (2008). Eight genes are required for functional reconstitution of the Caenorhabditis elegans levamisole-sensitive acetylcholine receptor. PNAS, 105(47), 18590-18595 .

Cancer and cell cycle regulation:

• Fang et al. (2015). UNC50 prompts G1/S transition and proliferation in HCC by regulation of epidermal growth factor receptor trafficking. PLOS ONE, 10(3), e0119338 .

• Selyunin et al. (2017). Genome-wide siRNA screen identifies UNC50 as a regulator of Shiga toxin 2 trafficking. Journal of Cell Biology, 216(10), 3249-3262 .