Anti-ACP5 monoclonal antibody, clone 8F7B22

Expression of selected genes in relation to phosphate (P-i) starvation and sugar sensing was studied in leaves of Arabidopsis. Excised leaf segments with different P status were supplied with combinations of P-i and sugars. Sugar-inducible genes, encoding beta-amylase (beta-AMY) and chalcone synthase (CHS), were also induced by P deficiency, and were more strongly regulated by sugars when leaf segments originated from P-starved plants. Furthermore, transcript levels of the P-starvation-inducible genes ACP5 (encoding an acid phosphatase), RNS1 (encoding a ribonuclease), and IPS1 (unknown function) increased in response to exogenously applied sugars. Supply of P-i to the leaf segments reversed both P-starvation-induced and sugar-induced gene expression. These interactions reveal a close relationship between P and sugar sensing. To differentiate between hexokinase-dependent and hexokinase-independent sugar sensing the effect of the glucose analogue 2-deoxyglucose and gene expression in the hexokinase-1 deficient mutant, gin2-1, were studied. Both beta-AMY and CHS were induced by supplying sucrose to excised leaves but not by 2-deoxyglucose, confirming that these genes are regulated by hexokinase-independent sugar sensing. In the gin2-1 mutant both beta-AMY and CHS responded clearly to P starvation excluding that hexokinase-1 mediates the response to P. Similarly, the P-responding genes, IPS1 and RNS1 were repressed by addition of P-i also in the gin2-1 mutant. In conclusion, several phosphate starvation-induced genes are also sugar-induced and hexokinase-independent sugar sensing in Arabidopsis is strongly intensified by phosphate starvation.

As a crucial virulence factor of Porphyromonas gingivalis, gingipains play an important role in periodontal destruction. This study aimed to investigate the effect of gingipains on osteoclastogenesis. We used RAW264.7 cells as osteoclast precursors in our study. In experimental groups, cells were treated with gingipains and/or receptor activator of nuclear factor-kappa B ligand (RANKL). Tartrate-resistant acid phosphatase (TRAP) activity staining assay showed osteoclast precursors and RANKL-induced mature osteoclasts were increased in a gingipains dose-dependent manner. Real-time reverse transcription polymerase chain reaction analysis demonstrated that gingipains upregulated osteoclastic genes including the protease cathepsin K (Ctsk), matrix metalloprotein 9 (Mmp9), nuclear factor of activated T cells 1 (Nfatc1) and acid phosphatase 5, tartrate resistant (Acp5) in a time-dependent manner. Western blotting assays presented upregulated expressions of TNF receptor-activating factor 6 (TRAF6) and integrin beta 3 induced by gingipains and RANKL compared to RANKL alone. Enhanced integrin-related signaling was also demonstrated by elevated phosphorylations of FAK and paxillin compared to control. Moreover, the pit resorption assays showed that gingipains augmented bone resorptive function of osteoclasts induced by RANKL. When we used Cilengitide to block integrin alpha v beta 3, gingipains reversed the reduction of formation and resorptive function in RANKL-induced osteoclasts, as they enhanced integrin alpha v beta 3 levels more than RANKL treatment alone. In conclusion, our data suggest that gingipains augmented the differentiation and function of mature osteoclasts induced by RANKL through the increase in integrin alpha v beta 3.