Poly(ADP-ribosyl)ation enhances HuR oligomerization and contributes to pro-inflammatory gene mRNA stabilization.

Poly(ADP-ribosyl)ation (PARylation) is an important post-translational modification mainly catalyzed by poly-ADP-ribose polymerase 1 (PARP1). In addition to having important roles in DNA damage detection and repair, it functions in gene expression regulation, especially at the posttranscriptional level. Embryonic lethal abnormal vision-like 1/human antigen R (ELAVL/HuR), a canonical 3' untranslated region AU-rich element-binding protein, is a crucial mRNA-stabilizing protein that protects target mRNAs from RNA-destabilizing protein- or microRNA-induced silencing complex (miRISC)-mediated degradation. Additionally, in some cases, HuR itself either promotes or suppresses translation. Here, we demonstrated that in response to inflammatory stimuli, the PARylation of HuR, mostly at the conserved D226 site, by PARP1 increased the formation of the HuR oligomer/multimer, and HuR oligomerization promoted the disassociation of miRISC and stabilized the pro-inflammatory gene mRNAs. The prevention of PARP1 activation or HuR oligomerization attenuated lipopolysaccharide-induced inflammatory gene expression and the airway recruitment of neutrophils in mouse lungs. The present study verified a novel mechanism of PARP1 and HuR PARylation in the RNA stability regulation, increasing our understanding of how PARP1 regulates gene expression.

Novel insights into PARPs in gene expression: regulation of RNA metabolism.

Poly(ADP-ribosyl)ation (PARylation) is an important post-translational modification in which an ADP-ribose group is transferred to the target protein by poly(ADP-riboses) polymerases (PARPs). Since the discovery of poly-ADP-ribose (PAR) 50 years ago, its roles in cellular processes have been extensively explored. Although research initially focused on the functions of PAR and PARPs in DNA damage detection and repair, our understanding of the roles of PARPs in various nuclear and cytoplasmic processes, particularly in gene expression, has increased significantly. In this review, we discuss the current advances in understanding the roles of PARylation with a particular emphasis in gene expression through RNA biogenesis and processing. In addition to updating PARP's significance in transcriptional regulation, we specifically focus on how PARPs and PARylation affect gene expression, especially inflammation-related genes, at the post-transcriptional levels by modulating RNA processing and degrading. Increasing evidence suggests that PARP inhibition is a promising treatment for inflammation-related diseases besides conventional chemotherapy for cancer.

Synergistic Inhibition of Enamel Demineralization by Peptide 8DSS and Fluoride.

The biomimetic peptide 8DSS has shown beneficial effects in promoting remineralization of demineralized enamel in vitro. Here we examined the ability of 8DSS alone and in combination with fluoride to inhibit enamel demineralization during pH-cycling mimicking intraoral conditions. Enamel blocks were subjected to 9 days of pH-cycling in the presence of 1,000 ppm NaF (positive control), distilled-deionized water (DDW; negative control), 25 µM 8DSS alone, 25 µM 8DSS with 500 ppm NaF (8DSS-FL) or 25 µM 8DSS with 1,000 ppm NaF (8DSS-FH) twice daily for 1 min each time. The blocks were analyzed in terms of surface microhardness (SMH), fluoride uptake and mineral content. The 8DSS-treated blocks showed significantly lower mineral loss, shallower lesions and higher SMH than the DDW-treated blocks. No significant differences were observed between the blocks treated with 8DSS alone or fluoride alone. The blocks treated with 8DSS alone or DDW showed similar amounts of fluoride uptake, which was the lowest of all the treatment groups. The blocks treated with 8DSS-FL or 8DSS-FH did not differ significantly, and both groups showed significantly greater SMH and fluoride uptake as well as significantly lower mineral loss and shallower lesions than the NaF-treated blocks. Mineral content was significantly higher in the 8DSS-treated blocks than in the DDW-treated blocks from the surface layer (10 µm) to the lesion depth (110 µm), and it was significantly higher in the blocks treated with 8DSS-FL or 8DSS-FH than in the NaF-treated blocks from 10 to 90 µm. These findings illustrate the potential of 8DSS for inhibiting enamel demineralization and for enhancing the anticaries effect of NaF.

Activity of Synthetic Antimicrobial Peptide GH12 against Oral Streptococci.

Controlling the growth of cariogenic microorganisms such as oral streptococci is an adjunct therapy for caries-active individuals to prevent and treat caries. Here we investigated the antimicrobial activity of the synthetic amphipathic α- helical antimicrobial peptide GH12 (GLLWHLLHHLLH-NH2) against oral streptococci in vitro. Circular dichroism studies showed that GH12 takes on an α-helical conformation in the presence of membrane-mimicking solvents, and reversed-phase high-performance liquid chromatography studies showed that GH12 remains stable in saliva. The peptide showed bactericidal activity against oral streptococci, with minimum inhibitory concentrations ranging from 6.7 to 32.0 µg/ml. GH12 concentrations 4-fold higher than the minimum bactericidal concentration completely killed oral streptococci within 20 min. Treating oral streptococci with GH12 caused noticeable changes in bacterial viability and morphology based on confocal laser scanning microscopy and scanning electron microscopy. Effects of GH12 on biofilm formation and on viability of mature biofilm were quantified by crystal violet staining and the 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide assay. GH12 effectively inhibited biofilm formation and metabolic activity in biofilms of oral streptococci, especially S. mutans, S. sobrinus and S. salivarius. These results suggest that GH12 shows rapid and strong antimicrobial activity against oral streptococci in vitro, opening the door to preclinical and clinical studies to explore its potential for caries prevention and treatment.

Promotion of enamel caries remineralization by an amelogenin-derived peptide in a rat model.

OBJECTIVE: An amelogenin-derived peptide has been shown to promote remineralization of demineralized enamel in an in vitro model of initial caries induced by pH cycling. The present study examines whether the peptide exerts similar effects within the complex oral environment in vivo. DESIGN: Specific pathogen-free Sprague-Dawley rats (n=36) were infected with Streptococcus mutans, given ad libitum access to Diet 2000 and drinking water supplemented with sucrose (10%, w/v), and then randomly divided into three groups treated with 25µM peptide solution, 1g/L NaF or deionized water. Molar teeth were swabbed twice daily with the respective solutions for 24days. Then animals were killed, their jaws were removed and caries lesions were analyzed using the quantitative light-induced fluorescence-digital (QLF-D) technique to measure changes in mineral content. To verify QLF-D results, caries were scored for lesion depth and size using the Keyes method, and analyzed using polarized light microscopy (PLM). RESULTS: Mineral gain was significantly higher in teeth treated with peptide or NaF than in teeth treated with water (p<0.05), based on the QLF-D results (ΔF and ΔQ). Incidence of smooth-surface and sulcal caries based on Keyes scores was similar in rats treated with peptide or NaF, and significantly lower in these groups than in rats treated with water (p<0.05). Lesions on teeth treated with peptide or NaF were shallower, based on PLM. No significant differences were observed between molar enamel caries treated with peptide or NaF. CONCLUSIONS: This amelogenin-derived peptide can promote remineralization in a rat caries model, indicating strong potential for clinical use.

Potential of an amelogenin based peptide in promoting reminerlization of initial enamel caries.

OBJECTIVE: In this study we give a preliminary study of a rationally designed small peptide, which is based on the enamel matrix protein amelogenin, to investigate its effect on remineralization of initial enamel caries lesions. DESIGN: A novel peptide was designed and synthesized to investigate its effects on the remineralization of initial enamel carious lesions during pH cycling that simulates intra-oral conditions. Initial lesions were created in bovine enamel blocks, which were then pH-cycled for 12 days in the presence of 25µM peptide, 1g/L NaF (positive control), 50mM HEPES buffer(negative control). Before and after pH cycling, enamel blocks were analyzed by surface microhardness testing, polarized light microscopy and transverse microradiography. RESULTS: Percentage of surface microhardness recovery (SMHR%) after pH cycling was significantly higher in peptide group than HEPES group. Lower lesion depth and less mineral mineral loss were found in peptide or NaF treatment groups after the cycling, and were significantly different to HEPES group. No significant differences were observed between the blocks treated with peptide and those treated with NaF. CONCLUSSION: This study provides in vitro evidence that this amelogenin based peptide promotes enamel caries remineralization, offering a promising remineralizing biomaterial in initial enamel carious treatment.