NUP210 Human

What is NUP210 and what is its primary function in the nuclear pore complex?

The most effective methods to study NUP210's structure and organization include:

• Immunofluorescence microscopy with NUP210-specific antibodies

• Cryo-electron microscopy of isolated nuclear pore complexes

• Co-immunoprecipitation studies to identify interaction partners

• CRISPR-Cas9 knockout models to assess structural dependencies

Is NUP210 expression universal across human tissues and cell types?

No, NUP210 shows tissue-specific expression patterns rather than universal expression. Unlike many core components of the nuclear pore complex, NUP210 is not ubiquitously expressed in all tissues . This differential expression suggests that NUP210 contributes to specialized functions of the nuclear pore complex in specific cell lineages.

Methodological approaches to characterize expression patterns include:

• Analysis of tissue-specific transcriptome databases such as GTEx

• Immunohistochemistry on tissue microarrays

• Western blotting of protein lysates from various tissues

• Single-cell RNA sequencing to identify cell type-specific expression

How is NUP210 implicated in acute myeloid leukemia (AML) pathogenesis?

NUP210 mRNA expression is significantly increased in the bone marrow of AML patients compared to healthy individuals . This upregulation correlates with specific AML subtypes according to the French-American-British (FAB) classification and is associated with bone marrow blast counts . Higher NUP210 expression serves as an independent biomarker of poor prognosis in AML (P < 0.05), with particularly strong prognostic significance in female patients but not male patients .

For researchers investigating NUP210 in AML, recommended methodological approaches include:

• Bioinformatic analysis of public databases (TCGA, GTEx)

• qPCR quantification of NUP210 expression in patient samples

• Survival analysis stratified by expression levels and patient characteristics

• Functional studies in AML cell lines with NUP210 knockdown/overexpression

What role does NUP210 play in breast cancer metastasis?

NUP210 has been identified as a metastasis susceptibility gene for human estrogen receptor positive (ER+) breast cancer . It functions as a cellular mechanosensor in this context. The mechanistic pathway involves:

• NUP210 interaction with LINC complex protein SUN2, connecting the nucleus to the cytoskeleton

• The NUP210/SUN2 complex interacting with chromatin via BRD4 and histones H3.1/H3.2 at the nuclear periphery

• In NUP210 knockout cells, mechanosensitive genes accumulate H3K27me3 heterochromatin modification

• This leads to transcriptional repression, resulting in defective mechanotransduction and focal adhesion

Experimental validation shows that NUP210 depletion suppresses lung metastasis in mouse models of breast cancer .

How can researchers develop NUP210-targeted therapeutic approaches?

When developing NUP210-targeted therapeutics, researchers should consider:

• Cell type specificity: Target NUP210 in cancer cells while preserving its function in normal tissues

• Sex-specific effects: Account for the differential impact of NUP210 in male versus female patients

• Mechanotransduction pathway: Target downstream effectors or upstream regulators of NUP210

• Combinatorial approaches: Use NUP210 status for patient stratification in combination with other therapies

Recommended experimental pipeline:

• High-throughput screening for small molecule inhibitors of NUP210-protein interactions

• CRISPR screens to identify synthetic lethal interactions with NUP210 depletion

• Validation in patient-derived xenograft models

• Assessment of effects on normal tissue function, particularly T cells

How does NUP210 deficiency affect T cell homeostasis?

NUP210 deficiency has a T cell-intrinsic impact on peripheral T cell homeostasis, particularly affecting CD4:CD8 T cell ratios . Interestingly, thymic development remains largely normal in NUP210-deficient mice, with no observed differences in double negative (DN), double positive (DP), or single positive (SP) thymocyte populations . Key transcription factors involved in CD4-CD8 lineage decisions (Tbx21, Runx3, and Thpok) remain unchanged throughout thymic differentiation .

Research approaches to investigate this phenotype include:

• Flow cytometry analysis of T cell subsets in various lymphoid tissues

• Mixed bone marrow chimeras to assess cell-intrinsic effects

• Competitive adoptive transfer experiments

• Transcriptomic profiling of affected T cell populations

• Analysis of T cell receptor signaling in NUP210-deficient cells

Why do NUP210 knockout mice remain viable despite the evolutionary conservation of this protein?

The viability of NUP210 knockout mice presents a fascinating biological paradox. Despite the deep evolutionary conservation of NUP210 across eukaryotes, mice deficient in this protein develop with no major phenotypes beyond altered T cell homeostasis . This surprising viability suggests:

• Functional redundancy: Other nucleoporins may compensate for NUP210 loss in most cell types

• Specialized functions: NUP210's essential roles may be restricted to specific contexts like T cells

• Stress response requirements: NUP210 might be dispensable under standard laboratory conditions but essential under specific stresses or challenges

This raises important questions about the evolutionary pressures maintaining NUP210 conservation. Researchers should consider:

• Comparative studies across species and cell types

• Challenge experiments exposing knockout mice to various stressors

• Deep phenotyping beyond standard health parameters

• Investigation of subtle cellular defects that might accumulate over time

What techniques are most effective for analyzing NUP210 interactions with chromatin architecture?

To analyze NUP210's interactions with chromatin and nuclear architecture, researchers should employ these techniques:

• Proximity-based approaches:

  • BioID or APEX2 proximity labeling to identify near-neighbors of NUP210

  • ChIP-seq to identify genomic regions associated with NUP210

  • DamID-seq as an antibody-independent approach to mapping interactions

• High-resolution imaging:

  • Super-resolution microscopy (STORM, PALM, SIM)

  • Correlative light and electron microscopy (CLEM)

  • Live-cell imaging with fluorescently tagged NUP210

• Chromatin conformation techniques:

  • Hi-C or derivatives (Micro-C, HiChIP) to map global chromatin organization

  • DNA FISH combined with immunofluorescence for locus-specific studies

  • ATAC-seq to assess chromatin accessibility changes upon NUP210 depletion

• Functional genomics:

  • CUT&RUN or CUT&Tag for more precise chromatin mapping

  • RNA-seq to correlate chromatin changes with gene expression

  • Epigenomic profiling focusing on H3K27me3 and other relevant modifications

How can researchers best study the mechanosensing function of NUP210?

To study NUP210's role as a mechanosensor, consider these methodological approaches:

• Controlled mechanical stimulation:

  • Atomic force microscopy to apply precise forces to the cell surface

  • Micropatterned substrates with varying stiffness

  • Fluid shear stress systems for endothelial models

  • Magnetic twisting cytometry for localized force application

• Real-time monitoring:

  • FRET-based tension sensors incorporated into NUP210 or interacting proteins

  • Live-cell imaging during mechanical stimulation

  • Calcium imaging to detect mechanosensitive signaling activation

• Molecular interaction studies:

  • Co-immunoprecipitation under different mechanical conditions

  • Proximity ligation assay to visualize protein interactions in situ

  • Mass spectrometry to identify mechano-responsive post-translational modifications

• Transcriptional readouts:

  • RNA-seq comparing wild-type and NUP210-deficient cells under mechanical stimulation

  • ChIP-seq for mechano-responsive transcription factors

  • Single-cell approaches to capture heterogeneous responses

What are the optimal approaches for studying sex-specific effects of NUP210?

To investigate the sex-specific effects of NUP210 observed in both AML patients and animal models, researchers should consider:

• Experimental design considerations:

  • Include both sexes in all experiments with sufficient power for sex-stratified analysis

  • Use gonadectomy models to distinguish hormonal from genetic sex effects

  • Consider four-core genotype mouse models to separate chromosomal sex from gonadal sex

• Hormone manipulation studies:

  • Assess NUP210 expression and function after hormone treatment

  • Use hormone receptor antagonists to block specific pathways

  • Test for hormone response elements in the NUP210 gene promoter and enhancers

• Molecular analyses:

  • Comparative transcriptomics of male vs. female cells with NUP210 perturbation

  • Epigenomic profiling to identify sex-specific chromatin landscapes

  • Proteomics to identify sex-specific NUP210 interactors

• Clinical correlations:

  • Analyze patient datasets with sex-stratified outcomes

  • Develop sex-specific biomarker thresholds for NUP210 expression

  • Correlate NUP210 effects with hormone receptor status in tumors

How might NUP210 be involved in cellular differentiation and development?

NUP210 appears to play critical roles in cellular differentiation processes. Previous studies have shown that shRNA knockdown of NUP210 in myoblasts and embryonic stem cells induces apoptosis and completely abrogates their differentiation into myotubes and neuroprogenitor cells . This suggests NUP210 functions as a scaffolding protein for transcriptional complexes such as Mef2C and contributes to cell fate determination through tissue-specific expression patterns .

Recommended research approaches include:

• Single-cell RNA-seq during differentiation processes

• Inducible knockout systems to assess temporal requirements

• ChIP-seq for lineage-specific transcription factors in NUP210-deficient cells

• Differentiation assays under varying mechanical conditions

• Rescue experiments with domain-specific NUP210 mutants

What is the relationship between NUP210 and cellular stress responses?

While not directly addressed in the provided search results, the relationship between NUP210 and cellular stress responses represents an important research direction. Based on its location at the nuclear pore and interaction with chromatin, NUP210 likely participates in stress response mechanisms through:

• Regulation of stress-responsive gene expression

• Modulation of nucleocytoplasmic transport of stress-related factors

• Integration of mechanical stress signals from the cytoskeleton

• Potential role in DNA damage responses at the nuclear periphery

Experimental approaches should include:

• Exposure of wild-type and NUP210-deficient cells to various stressors (oxidative, genotoxic, thermal, mechanical)

• Analysis of stress-responsive transcription factor localization and activity

• Assessment of DNA damage repair efficiency at peripheral vs. central nuclear regions

• Evaluation of nuclear envelope integrity under stress conditions

How can single-cell technologies advance our understanding of NUP210 function?

Single-cell technologies offer powerful approaches to dissect the heterogeneous functions of NUP210 across cell types and states:

• Single-cell RNA sequencing can:

  • Identify cell populations particularly dependent on NUP210

  • Reveal compensatory mechanisms in NUP210-deficient cells

  • Track transcriptional changes during differentiation or stress responses

• Single-cell proteomics/epitope profiling:

  • Map NUP210 protein levels across cell populations

  • Correlate with cell state markers and signaling pathways

  • Identify rare cell populations with unique NUP210 dependencies

• Single-cell chromatin accessibility and conformation:

  • Correlate nuclear architecture with NUP210 expression

  • Identify cell state-specific chromatin organization patterns

  • Link gene positioning to transcriptional outcomes

• Spatial transcriptomics:

  • Map NUP210-dependent gene expression in tissue contexts

  • Correlate with microenvironmental factors and mechanical forces

  • Identify niche-specific requirements for NUP210 function