NAA50 Human

What is human NAA50 and what is its basic cellular function?

Human N-alpha-acetyltransferase 50 (NAA50) is a 169 amino acid cytoplasmic protein belonging to the acetyltransferase family and GNAT subfamily . It functions as a catalytic component of the ARD1A-NARG1 complex, displaying alpha acetyltransferase activity . NAA50 is encoded by a gene mapping to human chromosome 3q13.2 .
Methodologically, NAA50 can be studied as part of the NatE complex, which consists of Naa10, Naa50, and the auxiliary subunit Naa15 . It plays a critical role in co-translational modification of newly synthesized proteins in eukaryotic cells . Specifically, human Naa50 acetylates methionine followed by any residue except for proline, indicating its broad substrate specificity .

Where is NAA50 protein localized in human cells and how can this be determined?

NAA50 protein is primarily localized to the nucleoli in human cells, as demonstrated through immunofluorescence studies . Research using the Human Protein Atlas database has shown that NAA50 protein can be visualized in the nucleoli of A-431 and U-2 OS cells using specific antibody staining techniques .
For visualization purposes, DAPI staining is used for nuclear localization (shown in blue), while NAA50 protein staining appears in green using specific antibodies . Interestingly, while subcellular localization studies show nucleolar concentration, immunohistochemistry results across cancer tissues demonstrate moderate to strong cytoplasmic staining, suggesting potential context-dependent localization patterns .

How can researchers express and purify recombinant human NAA50 for functional studies?

For experimental studies requiring purified NAA50, researchers can express recombinant human NAA50 protein with an N-terminal His-tag in E. coli expression systems . Purification is typically accomplished using conventional chromatography techniques, with product validation performed via SDS-PAGE to confirm protein identity and purity .
The recombinant protein can be used for various applications including enzymatic assays, crystallization studies, and protein-protein interaction analyses. When designing expression constructs, researchers should consider that human NAA50 consists of 169 amino acids, and the addition of affinity tags may affect certain functional properties .

How does NAA50 expression vary across different cancer types and what methodologies best capture this variation?

Pan-cancer analysis reveals differential NAA50 expression patterns across cancer types, with significant overexpression in multiple cancers compared to normal tissues. Studies using the TIMER2.0 database demonstrated increased NAA50 expression in BLCA, CESC, COAD, ESCA, HNSC, LIHC, LUAD, LUSC, and STAD .
Additional analysis through SangerBox3.0 database of TCGA and GTEx datasets confirmed and expanded these findings to include GBM, GBMLGG, LGG, UCEC, BRCA, and numerous other cancer types . Interestingly, NAA50 was found to be downregulated in kidney-related cancers including KICH, KIRC, KIRP, and PCPG, suggesting tissue-specific regulatory mechanisms .
For comprehensive pan-cancer analysis, researchers should employ multiple database tools including TIMER2.0, SangerBox3.0, and GEPIA, along with validation through protein-level analysis using immunohistochemistry via resources like the Human Protein Atlas .

How does NAA50 influence cancer cell proliferation and what experimental approaches can measure this effect?

NAA50 significantly impacts cancer cell proliferation, particularly in lung adenocarcinoma. In vitro experiments using knockout models demonstrate that silencing NAA50 significantly inhibits proliferation rates in H1299 and PC9 lung adenocarcinoma cell lines .
For experimental validation, researchers can employ multiple complementary approaches:

• Western blot analysis to confirm NAA50 knockdown efficiency

• CCK-8 assay to evaluate cell proliferation rates over time

• EdU assay to directly visualize and quantify cellular proliferation
  These combined approaches have demonstrated that NAA50 downregulation significantly inhibits proliferation in lung adenocarcinoma cells, confirming computational predictions that NAA50 is involved in cell cycle regulation and proliferation .

What is the relationship between NAA50 expression and immune cell infiltration in cancer?

NAA50 expression shows significant correlations with immune cell infiltration patterns across cancer types. Analysis using the TIMER2.0 database reveals that NAA50 expression is:

• Positively correlated with bone myeloid-derived suppressor cell (MDSC) infiltration in multiple cancer types

• Negatively correlated with natural killer T (NKT) cell infiltration across various cancers

• Associated with altered infiltration levels of B cells, CD4+ T cells, CD8+ T cells, macrophages, and neutrophils specifically in LUAD
  Additionally, correlation analysis between NAA50 expression and major histocompatibility complexes (MHCs) and chemokines using the TISIDB database provides further insights into NAA50's role in tumor immune microenvironment regulation .
  These findings suggest NAA50 may influence tumor immune evasion mechanisms, making it a potential target for immunotherapy approaches.

How can single-cell sequencing data be utilized to understand NAA50's functional role in cancer?

Single-cell sequencing provides powerful insights into NAA50's cancer-related functions. Analysis using CancerSEA's single-cell sequence data reveals that NAA50 positively correlates with cell cycle and invasion processes in most tumors, with particularly prominent effects in LUAD .
Methodologically, researchers can leverage single-cell data to:

• Identify co-expressed genes (top 50 positively and negatively correlated genes)

• Perform GO analysis to identify biological functions (revealing NAA50's involvement in chromosome segregation, DNA replication, cell cycle phase transition, etc.)

• Conduct KEGG pathway analysis showing enrichment in cell cycle, RNA transport, DNA replication, Fanconi anemia pathway, p53 signaling, and other critical pathways
  This multi-layered analysis approach helps elucidate NAA50's mechanistic role in cancer progression, particularly its function in cell cycle regulation.

What is the relationship between NAA50 and DNA methylation in cancer?

DNA methylation analysis of NAA50 provides important insights into its regulation in cancer. Research shows that the methylation level of NAA50 promoter is higher in ESCA, KIRC, LIHC, LUSC, PAAD, and SARC compared to adjacent normal tissues, while decreased in BRCA and HNSC .
This cancer-specific methylation pattern may explain differential expression levels and contribute to NAA50's role in tumorigenesis. DNA methylation involves the production of 5-methylcytosine by adding a methyl group to cytosine under DNMT catalysis .
For comprehensive understanding, researchers should consider that DNA methylation not only serves as an important marker for drug efficacy, survival rates, and cancer recurrence, but is also closely related to immune responses in various cancers . Integrating methylation analysis with expression and functional data provides a more complete picture of NAA50's role in cancer biology.

What are effective approaches for studying NAA50 knockdown effects in cancer models?

When designing experiments to study NAA50 knockdown effects, researchers should consider multiple technical approaches:

• siRNA or shRNA approaches: For transient or stable knockdown of NAA50 in cell lines

• CRISPR-Cas9 system: For complete knockout studies or precise genetic modifications

• Validation of knockdown efficiency: Western blot analysis is essential to confirm reduced NAA50 protein levels

• Selection of appropriate cell lines: Based on search results, H1299 and PC9 lung adenocarcinoma cell lines have been successfully used to study NAA50 function

• Multiple functional assays: Employ CCK-8 for proliferation rate assessment and EdU assay for direct visualization of cellular proliferation
  This comprehensive experimental design allows for robust assessment of NAA50's functional roles in cancer biological processes.

How can researchers effectively analyze the relationship between NAA50 and cell cycle regulation?

To investigate NAA50's role in cell cycle regulation, researchers should employ a multi-faceted approach:

• Bioinformatic analysis: Examine correlations between NAA50 expression and CDK family, CDKI family, and Cyclin family genes using databases like TIMER2.0 (e.g., correlation with CDK1)

• GO and KEGG pathway analysis: Identify enrichment in cell cycle-related pathways and biological processes, including chromosome segregation, mitotic cell cycle phase transition, and cell cycle checkpoints

• Flow cytometry: For cell cycle profile analysis following NAA50 manipulation

• Western blot analysis: Evaluate expression changes in key cell cycle regulators (cyclins, CDKs) after NAA50 knockdown

• Single-cell analysis: Correlate NAA50 expression with cell cycle states across different cancer types
  This integrated approach provides comprehensive insights into NAA50's mechanistic role in cell cycle regulation across cancer contexts.

What databases and computational tools are most valuable for comprehensive NAA50 research?

For comprehensive NAA50 research, investigators should utilize multiple specialized databases and computational tools:

How might NAA50 serve as a biomarker for cancer diagnosis or prognosis?

NAA50 shows considerable potential as a cancer biomarker based on several consistent findings:

What is the potential for NAA50 as a therapeutic target in cancer treatment?

NAA50's characteristics suggest significant potential as a therapeutic target:

• Functional importance: NAA50 knockout significantly inhibits cancer cell proliferation, particularly in LUAD models

• Pathway involvement: NAA50 regulates critical pathways including cell cycle, DNA replication, and p53 signaling

• Immune correlation: NAA50 expression correlates with immune cell infiltration patterns, suggesting potential combination with immunotherapy approaches

• Cancer-specific overexpression: Elevated in most cancer types while showing lower expression in normal tissues, potentially offering a therapeutic window
  Therapeutic strategies might include development of small molecule inhibitors targeting NAA50's acetyltransferase activity, siRNA-based approaches for expression suppression, or targeting NAA50-dependent pathways. Further research should focus on identifying cancer types most dependent on NAA50 function and potential resistance mechanisms.

What are the key unresolved questions regarding NAA50's role in human biology and disease?

Several critical questions remain unanswered regarding NAA50:

• Substrate specificity mechanism: While we know NAA50 acetylates methionine followed by any residue except proline , the structural basis for this specificity remains to be fully characterized

• Tissue-specific regulation: The mechanisms explaining why NAA50 is downregulated in kidney cancers while upregulated in most other cancer types need investigation

• Interaction with immune system: How NAA50 influences immune cell infiltration and potentially contributes to immune evasion needs deeper exploration

• Post-translational modifications: How NAA50 itself is regulated through potential phosphorylation, ubiquitination, or other modifications remains largely unknown

• Therapeutic resistance: Potential mechanisms of resistance to NAA50-targeted therapies need to be anticipated and studied
  Addressing these questions would significantly advance our understanding of NAA50 biology and its therapeutic potential.

How can multi-omics approaches enhance our understanding of NAA50 function in human diseases?

Integration of multiple omics technologies offers comprehensive insights into NAA50 function:

• Genomics: Further analysis of NAA50 genetic variations, copy number alterations, and promoter variants across populations and disease states

• Transcriptomics: RNA-seq and single-cell transcriptomics to understand cell-type specific expression patterns and regulatory networks

• Proteomics: Mass spectrometry-based approaches to identify the complete set of proteins acetylated by NAA50 and how this acetylome changes in disease states

• Epigenomics: Deeper investigation of NAA50 promoter methylation patterns and their functional consequences in different cancer types

• Metabolomics: Investigation of how NAA50 activity impacts cellular metabolism, particularly in cancer contexts This integrated approach would provide a systems-level understanding of NAA50's role in human biology and disease, potentially revealing novel therapeutic opportunities and biomarker applications.