NAT10-mediated acetylation of JunB drives melanoma progression by enforcing SLC27A1-dependent lipid metabolic reprogramming

Background Aberrant protein acetylation homeostasis participates in regulating tumor cell proliferation, metabolic reprogramming, and genomic stability. Nonetheless, it is uncertain whether and how protein acetylation modification patterns affect clinical outcomes in melanoma patients. Methods Single-cell RNA sequencing and transcriptomic data, combined with machine learning, were used to identify N-acetyltransferase 10 (NAT10) as a key gene driving MM progression. NAT10 expression and its prognostic value were evaluated by immunohistochemistry on tissue microarrays and validated by quantitative real-time PCR and Western blot. Assay for Transposase-Accessible Chromatin using sequencing was performed to assess NAT10-mediated changes in chromatin accessibility. TurboID proximity labeling coupled with mass spectrometry was used to identify NAT10-interacting proteins and acetylation sites on JunB. Co immunoprecipitation experiments, immunofluorescence experiments, and molecular docking were performed to detect the interaction between NAT10 and JunB, as well as its regulation of JunB acetylation and ubiquitination modification. Cleavage Under Targets and Tagmentation, Chromatin immunoprecipitation, and dual-luciferase reporter assays were used to elucidate the mechanism by which NAT10 promotes solute carrier family 27 member 1 (SLC27A1) transcription via JunB. Non-targeted metabolomics combined with in vitro and in vivo functional assays assessed the effects of NAT10, JunB, and SLC27A1 on melanoma malignant phenotypes. Results We identify NAT10 as a previously unrecognized prognostic acetyltransferase that is aberrantly upregulated in melanoma, where it promotes tumor stemness and metastasis in correlation with poor clinical outcome. Mechanistically, NAT10 directly acetylates JunB at lysine 343 (K343), which competitively inhibits K48-linked ubiquitination and subsequent proteasomal degradation of JunB, thereby enhancing its stability and transcriptional activity. Acetylated JunB subsequently transactivates SLC27A1, driving lipid metabolic reprogramming and fuels melanoma proliferation, stemness, invasion, and metastasis. Importantly, combined administration of the NAT10 inhibitor Remodelin and the SLC27A1 inhibitor FATP1-IN-1 exerts synergistic antitumor and anti-metastatic effects in vivo, significantly outperforming either monotherapy. Conclusion This study identified NAT10 as an oncogenic driver that promotes melanoma progression and metastasis via the NAT10/JunB/SLC27A1 axis, a previously unrecognized pathway governing lipid metabolic reprogramming. Moreover, concurrent pharmacological inhibition of NAT10 and SLC27A1 produces synergistic antitumor effects in vivo, establishing a promising therapeutic strategy with high translational potential for melanoma treatment.