Sterol regulatory element-binding proteins (SREBPs) are the key transcription factors that modulate lipid biosynthesis. SREBPs are synthesized as endoplasmic reticulum-bound precursors that require proteolytic activation in the Golgi apparatus. The stability and maturation of precursor SREBPs depend on their binding to SREBP cleavage-activating protein (SCAP), which escorts the SCAP-SREBP complex to the Golgi apparatus. In this study, we identified heat shock protein (HSP) 90 as a novel SREBP regulator that binds to and stabilizes SCAP-SREBP. In HepG2 cells, HSP90 inhibition led to proteasome-dependent degradation of SCAP-SREBP, which resulted in the down-regulation of SREBP target genes and the reduction in intracellular triglyceride and cholesterol levels. We also demonstrated in vivo that HSP90 inhibition decreased SCAP-SREBP protein, down-regulated SREBP target genes, and reduced lipids levels in mouse livers. We propose that HSP90 plays an indispensable role in SREBP regulation by stabilizing the SCAP-SREBP complex, facilitating the activation of SREBP to maintain lipids homeostasis.

Sterol regulatory element–binding protein 2 (SREBP2) is the master transcription factor that regulates cholesterol metabolism. SREBP2 activation is regulated by SREBP chaperone SCAP. Here we show that ring finger protein 5 (RNF5), an endoplasmic reticulum-anchored E3 ubiquitin ligase, mediates the Lys-29–linked polyubiquitination of SCAP and thereby activates SREBP2. RNF5 knockdown inhibited SREBP2 activation and reduced cholesterol biosynthesis in human hepatoma cells, and RNF5 overexpression activated SREBP2. Mechanistic studies revealed that RNF5 binds to the transmembrane domain of SCAP and ubiquitinates the Lys-305 located in cytosolic loop 2 of SCAP. Moreover, the RNF5-mediated ubiquitination enhanced an interaction between SCAP luminal loop 1 and loop 7, a crucial event for SREBP2 activation. Notably, an overexpressed K305R SCAP variant failed to restore the SREBP2 pathway in SCAP-deficient cell lines. These findings define a new mechanism by which an ubiquitination-induced SCAP conformational change regulates cholesterol biosynthesis.

Anthracnose caused by Colletotrichum leads to a tremendous post-harvest mango loss. While chemical fungicides are applied to control anthracnose, natural alternatives are preferred due to food safety and environmental concerns. Pomelo extract (PE) exhibits a broad spectrum of antimicrobial activities; however, its effect against anthracnose is unknown. Here we investigated the chemical profile of PE using GC-MS and the anti-anthracnose activity of PE using in vitro and in vivo assays. We also evaluated the impact of storage temperature (0°, 5°, 10°, 20°, −20°, and −80 °C) and light conditions on the composition and antifungal activity of PE. We found that PE inhibited C. gloeosporioides in vitro with an IC₅₀ of 3.2 mL L⁻¹. Applying chitosan-based coating incorporated with 20 mL L⁻¹ PE significantly suppressed anthracnose in post-harvest ‘Keitt’ mango. A storage temperature below 5 °C substantially preserved major compounds and the antifungal activity of PE after 6 m of storage. Finally, we showed that applying d-limonene, the key constituent of PE, inhibited C. gloeosporioides in vitro (IC₅₀: 10.9 mM) and suppressed anthracnose in vivo. In conclusion, we demonstrated that the application of PE and d-limonene are sustainable methods for anthracnose control in post-harvest crops and established the preservation protocol for PE.