Arabidopsis protein S-acyl transferases positively mediate BR signaling through S-acylation of BSK1.

Protein S-acyl transferases (PATs) catalyze S-acylation, a reversible post-translational modification critical for membrane association, trafficking, and stability of substrate proteins. Many plant proteins are potentially S-acylated but few have corresponding PATs identified. By using genomic editing, confocal imaging, pharmacological, genetic, and biochemical assays, we demonstrate that three Arabidopsis class C PATs positively regulate BR signaling through S-acylation of BRASSINOSTEROID-SIGNALING KINASE1 (BSK1). PAT19, PAT20, and PAT22 associate with the plasma membrane (PM) and the trans-Golgi network/early endosome (TGN/EE). Functional loss of all three genes results in a plethora of defects, indicative of reduced BR signaling and rescued by enhanced BR signaling. PAT19, PAT20, and PAT22 interact with BSK1 and are critical for the S-acylation of BSK1, and for BR signaling. The PM abundance of BSK1 was reduced by functional loss of PAT19, PAT20, and PAT22 whereas abolished by its S-acylation-deficient point mutations, suggesting a key role of S-acylation in its PM targeting. Finally, an active BR analog induces vacuolar trafficking and degradation of PAT19, PAT20, or PAT22, suggesting that the S-acylation of BSK1 by the three PATs serves as a negative feedback module in BR signaling.

The effect of vascular endothelial growth factor (VEGF) on mouse condylar articular cartilage cultured in vitro.

In this study, we explored the direct effect of vascular endothelial growth factor (VEGF) on temporomandibular joint osteoarthritis (TMJ-OA) by analyzing the transformation of mouse condylar cartilage treated in vitro with various concentrations of VEGF. Tissue samples from 126 condyles of four-week-old male C57 mice were randomly divided into 21 groups and treated with VEGF (0 ng/mL, 100 ng/mL, 500 ng/mL, 1 µg/mL, or 2 µg/mL). Furthermore, the samples were treated at different time points (1 d, 2 d, 4 d, and 7 d) and stained with hematoxylin and eosin (HE) and Safranin O and Fast Green stains to observe their morphology. The Mankin score was used to evaluate changes to the condylar cartilage tissues, and immunohistochemistry was performed to observe the expressions of VEGF receptor 2 (VEGFR2), matrix metallopeptidase 9 (MMP9), matrix metallopeptidase 13 (MMP13), and tumor necrosis factor-related activation-induced cytokine (TRANCE). An HE staining analysis revealed that the experimental groups treated with VEGF exhibited the destruction of their condylar cartilage and a proliferation of their hypertrophic cells, in comparison to the control group. Safranin O and Fast Green staining showed that the experimental groups had decreased levels of proteoglycan and degenerative changes in their condylar cartilage. The Mankin score of the samples increased with increasing concentration and treatment time of VEGF, and the differences between the groups were statistically significant (P < 0.05). Immunohistochemistry demonstrated that the expression levels of VEGFR2, MMP9, MMP13, and TRANCE significantly increased in the experimental groups, in comparison to those in the control group, suggesting that VEGF promoted TMJ-OA in mice in vitro.