RNA polymerase I subunit RPA43 activates rRNA expression and cell proliferation but inhibits cell migration.

The products synthesized by RNA polymerase I (Pol I) play fundamental roles in several cellular processes, including ribosomal biogenesis, protein synthesis, cell metabolism, and growth. Deregulation of Pol I products can cause various diseases such as ribosomopathies, leukaemia, and solid tumours. However, the detailed mechanism of Pol I-directed transcription remains elusive, and the roles of Pol I subunits in rRNA synthesis and cellular activities still need clarification. In this study, we found that RPA43 expression levels positively correlate with Pol I product accumulation and cell proliferation, indicating that RPA43 activates these processes. Unexpectedly, RPA43 depletion promoted HeLa cell migration, suggesting that RPA43 functions as a negative regulator in cell migration. Mechanistically, RPA43 positively modulates the recruitment of Pol I transcription machinery factors to the rDNA promoter by activating the transcription of the genes encoding Pol I transcription machinery factors. RPA43 inhibits cell migration by dampening the expression of c-JUN and Integrin. Collectively, we found that RPA43 plays opposite roles in cell proliferation and migration except for driving Pol I-dependent transcription. These findings provide novel insights into the regulatory mechanism of Pol I-mediated transcription and cell proliferation and a potential pathway to developing anti-cancer drugs using RPA43 as a target.

STAT3 promotes RNA polymerase III-directed transcription by controlling the miR-106a-5p/TP73 axis.

Deregulation of Pol III products causes a range of diseases, including neural diseases and cancers. However, the factors and mechanisms that modulate Pol III-directed transcription remain to be found, although massive advances have been achieved. Here, we show that STAT3 positively regulates the activities of Pol III-dependent transcription and cancer cell growth. RNA-seq analysis revealed that STAT3 inhibits the expression of TP73, a member of the p53 family. We found that TP73 is not only required for the regulation of Pol III-directed transcription mediated by STAT3 but also independently suppresses the synthesis of Pol III products. Mechanistically, TP73 can disrupt the assembly of TFIIIB subunits and inhibit their occupancies at Pol III target loci by interacting with TFIIIB subunit TBP. MiR-106a-5p can activate Pol III-directed transcription by targeting the TP73 mRNA 3' UTR to reduce TP 73 expression. We show that STAT3 activates the expression of miR-106a-5p by binding to the miRNA promoter, indicating that the miR-106a-5p links STAT3 with TP73 to regulate Pol III-directed transcription. Collectively, these findings indicate that STAT3 functions as a positive regulator in Pol III-directed transcription by controlling the miR-106a-5p/TP73 axis.

Transcription factor AP-2α activates RNA polymerase III-directed transcription and tumor cell proliferation by controlling expression of c-MYC and p53.

Deregulation of transcription factor AP2 alpha (TFAP2A) and RNA polymerase III (Pol III) products is associated with tumorigenesis. However, the mechanism underlying this event is not fully understood and the connection between TFAP2A and Pol III-directed transcription has not been investigated. Here, we report that TFAP2A functions as a positive factor in the regulation of Pol III-directed transcription and cell proliferation. We found TFAP2A is also required for the activation of Pol III transcription induced by the silencing of filamin A, a well-known cytoskeletal protein and an inhibitor in Pol III-dependent transcription identified previously. Using a chromatin immunoprecipitation technique, we showed TFAP2A positively modulates the assembly of Pol III transcription machinery factors at Pol III-transcribed gene loci. We found TFAP2A can activate the expression of Pol III transcription-related factors, including BRF1, GTF3C2, and c-MYC. Furthermore, we demonstrate TFAP2A enhances expression of MDM2, a negative regulator of tumor suppressor p53, and also inhibits p53 expression. Finally, we found MDM2 overexpression can rescue the inhibition of Pol III-directed transcription and cell proliferation caused by TFAP2A silencing. In summary, we identified that TFAP2A can activate Pol III-directed transcription by controlling multiple pathways, including general transcription factors, c-MYC and MDM2/p53. The findings from this study provide novel insights into the regulatory mechanisms of Pol III-dependent transcription and cancer cell proliferation.

TFIIB-related factor 1 is a nucleolar protein that promotes RNA polymerase I-directed transcription and tumour cell growth.

Eukaryotic RNA polymerase I (Pol I) products play fundamental roles in ribosomal assembly, protein synthesis, metabolism and cell growth. Abnormal expression of both Pol I transcription-related factors and Pol I products causes a range of diseases, including ribosomopathies and cancers. However, the factors and mechanisms governing Pol I-dependent transcription remain to be elucidated. Here, we report that transcription factor IIB-related factor 1 (BRF1), a subunit of transcription factor IIIB required for RNA polymerase III (Pol III)-mediated transcription, is a nucleolar protein and modulates Pol I-mediated transcription. We showed that BRF1 can be localized to the nucleolus in several human cell types. BRF1 expression correlates positively with Pol I product levels and tumour cell growth in vitro and in vivo. Pol III transcription inhibition assays confirmed that BRF1 modulates Pol I-directed transcription in an independent manner rather than through a Pol III product-to-45S pre-rRNA feedback mode. Mechanistically, BRF1 binds to the Pol I transcription machinery components and can be recruited to the rDNA promoter along with them. Additionally, alteration of BRF1 expression affects the recruitment of Pol I transcription machinery components to the rDNA promoter and the expression of TBP and TAF1A. These findings indicate that BRF1 modulates Pol I-directed transcription by controlling the expression of selective factor 1 subunits. In summary, we identified a novel role of BRF1 in Pol I-directed transcription, suggesting that BRF1 can independently regulate both Pol I- and Pol III-mediated transcription and act as a key coordinator of Pol I and Pol III.

STAT3 potentiates RNA polymerase I-directed transcription and tumor growth by activating RPA34 expression.

BACKGROUND: Deregulation of either RNA polymerase I (Pol I)-directed transcription or expression of signal transducer and activator of transcription 3 (STAT3) correlates closely with tumorigenesis. However, the connection between STAT3 and Pol I-directed transcription hasn't been investigated. METHODS: The role of STAT3 in Pol I-directed transcription was determined using combined techniques. The regulation of tumor cell growth mediated by STAT3 and Pol I products was analyzed in vitro and in vivo. RNAseq, ChIP assays and rescue assays were used to uncover the mechanism of Pol I transcription mediated by STAT3. RESULTS: STAT3 expression positively correlates with Pol I product levels and cancer cell growth. The inhibition of STAT3 or Pol I products suppresses cell growth. Mechanistically, STAT3 activates Pol I-directed transcription by enhancing the recruitment of the Pol I transcription machinery to the rDNA promoter. STAT3 directly activates Rpa34 gene transcription by binding to the RPA34 promoter, which enhances the occupancies of the Pol II transcription machinery factors at this promoter. Cancer patients with RPA34 high expression lead to poor survival probability and short survival time. CONCLUSION: STAT3 potentiates Pol I-dependent transcription and tumor cell growth by activating RPA34 in vitro and in vivo.

Early Growth Response 1 Strengthens Pol-III-Directed Transcription and Transformed Cell Proliferation by Controlling PTEN/AKT Signalling Activity.

RNA polymerase III (Pol III) products play essential roles in ribosome assembly, protein synthesis, and cell survival. Deregulation of Pol-III-directed transcription is closely associated with tumorigenesis. However, the regulatory pathways or factors controlling Pol-III-directed transcription remain to be investigated. In this study, we identified a novel role of EGR1 in Pol-III-directed transcription. We found that Filamin A (FLNA) silencing stimulated EGR1 expression at both RNA and protein levels. EGR1 expression positively correlated with Pol III product levels and cell proliferation activity. Mechanistically, EGR1 downregulation dampened the occupancies of Pol III transcription machinery factors at the loci of Pol III target genes. Alteration of EGR1 expression did not affect the expression of p53, c-MYC, and Pol III general transcription factors. Instead, EGR1 activated RhoA expression and inhibited PTEN expression in several transformed cell lines. We found that PTEN silencing, rather than RhoA overexpression, could reverse the inhibition of Pol-III-dependent transcription and cell proliferation caused by EGR1 downregulation. EGR1 could positively regulate AKT phosphorylation levels and is required for the inhibition of Pol-III-directed transcription mediated by FLNA. The findings from this study indicate that EGR1 can promote Pol-III-directed transcription and cell proliferation by controlling the PTEN/AKT signalling pathway.

Combating biofilms of foodborne pathogens with bacteriocins by lactic acid bacteria in the food industry.

Most foodborne pathogens have biofilm-forming capacity and prefer to grow in the form of biofilms. Presence of biofilms on food contact surfaces can lead to persistence of pathogens and the recurrent cross-contamination of food products, resulting in serious problems associated with food safety and economic losses. Resistance of biofilm cells to conventional sanitizers urges the development of natural alternatives to effectively inhibit biofilm formation and eradicate preformed biofilms. Lactic acid bacteria (LAB) produce bacteriocins which are ribosomally synthesized antimicrobial peptides, providing a great source of nature antimicrobials with the advantages of green and safe properties. Studies on biofilm control by newly identified bacteriocins are increasing, targeting primarily onListeria monocytogenes, Staphylococcus aureus, Salmonella, and Escherichia coli. This review systematically complies and assesses the antibiofilm property of LAB bacteriocins in controlling foodborne bacterial-biofilms on food contact surfaces. The bacteriocin-producing LAB genera/species, test method (inhibition and eradication), activity spectrum and surfaces are discussed, and the antibiofilm mechanisms are also argued. The findings indicate that bacteriocins can effectively inhibit biofilm formation in a dose-dependent manner, but are difficult to disrupt preformed biofilms. Synergistic combination with other antimicrobials, incorporation in nanoconjugates and implementation of bioengineering can help to strengthen their antibiofilm activity. This review provides an overview of the potential and application of LAB bacteriocins in combating bacterial biofilms in food processing environments, assisting in the development and widespread use of bacteriocin as a promising antibiofilm-agent in food industries.

Transcription factor GATA4 drives RNA polymerase III-directed transcription and transformed cell proliferation through a filamin A/GATA4/SP1 pathway.

RNA polymerase III (pol III) products play fundamental roles in a variety of cellular processes, including protein synthesis and cancer cell proliferation. In addition, dysregulation of pol III-directed transcription closely correlates with tumorigenesis. It is therefore of interest to identify novel pathways or factors governing pol III-directed transcription. Here, we show that transcription factor (TF) GATA binding protein 4 (GATA4) expression in SaOS2 cells was stimulated by the silencing of filamin A (FLNA), a repressor of pol III-directed transcription, suggesting that GATA4 is potentially associated with the regulation of pol III-directed transcription. Indeed, we show that GATA4 expression positively correlates with pol III-mediated transcription and tumor cell proliferation. Mechanistically, we found that GATA4 depletion inhibits the occupancies of the pol III transcription machinery factors at the loci of pol III target genes by reducing expression of both TFIIIB subunit TFIIB-related factor 1 and TFIIIC subunit general transcription factor 3C subunit 2 (GTF3C2). GATA4 has been shown to activate specificity factor 1 (Sp1) gene transcription by binding to the Sp1 gene promoter, and Sp1 has been confirmed to activate pol III gene transcription by directly binding to both Brf1 and Gtf3c2 gene promoters. Thus, the findings from this study suggest that GATA4 links FLNA and Sp1 signaling to form an FLNA/GATA4/Sp1 axis to modulate pol III-directed transcription and transformed cell proliferation. Taken together, these results provide novel insights into the regulatory mechanism of pol III-directed transcription.

Strategies to enhance micropollutant removal from wastewater by membrane bioreactors: Recent advances and future perspectives.

Membrane bioreactor (MBR) has been widely implemented to advance wastewater treatment and reuse. Nevertheless, conventional MBRs with porous microfiltration or ultrafiltration membranes are not designed for the removal of micropollutants, which ubiquitously occur in wastewater at trace concentrations, but potentially exert detrimental impacts to the ecosystem. Several effective strategies have been applied to improve MBR performance for micropollutant removal, particularly the hydrophilic and recalcitrant compounds. These strategies mainly include the optimization of operational conditions, employment of high-retention membranes to replace porous ones, addition of functional materials into bioreactor, and integration of effluent purification processes. In particular, effluent purification by advanced oxidation processes (AOPs) and high-retention membranes can complement MBR to secure almost complete removal of micropollutants. Nevertheless, further research is still necessary to evaluate the technical and economic feasibility of these strategies, especially for long-term treatment performance, to screen the suitable techniques for industrial MBR applications.

RNA polymerase I subunit 12 plays opposite roles in cell proliferation and migration.

RNA polymerase I (Pol I) is responsible for the synthesis of the majority of ribosomal RNA molecules in eukaryotes. Pol I subunit 12 (RPA12) is involved in the transcriptional termination and lipid metabolism in yeast. However, its role in human cells hasn't been investigated so far. Here, we show that RPA12 is present in the nucleolus and nucleoplasm of HeLa cells. RPA12 can act as a positive factor to regulate Pol I-mediated transcription and the proliferation of 293T and HeLa cells. Unexpectedly, RPA12 can repress HeLa cell migration, indicating that RPA12 plays opposite roles in cell proliferation and migration. This study provides a novel insight into the role of RPA12 in human cells.

Removal of antibiotics by sequencing-batch membrane bioreactor for swine wastewater treatment.

This study investigated the removal of antibiotics by sequencing-batch membrane bioreactor (SMBR) for swine wastewater treatment. Nine compounds categorized into three groups of commonly used veterinary antibiotics, namely sulfonamides, tetracyclines and fluoroquinolones, were evaluated. Results showed that both sulfonamides and tetracyclines were efficiently removed by SMBR (>90%) while a lower removal was observed for fluoroquinolones (<70%). Mass balance analysis evidenced that biodegradation/biotransformation was the main mechanism for the removal of antibiotics in SMBR operation. Moreover, sludge adsorption and membrane retention also slightly contributed to antibiotic removal. Of the three groups of antibiotics, tetracyclines and fluoroquinolones were more prone to accumulate in biosolids. It is noteworthy that antibiotics temporarily affected SMBR performance by inhibiting sludge growth and activity as well as increasing the concentrations of extracellular polymeric substances and soluble microbial products in the mixed liquor. Nevertheless, >60% of organic matter and nutrients in swine wastewater could be removed over SMBR operation.

Resource recovery from wastewater by anaerobic membrane bioreactors: Opportunities and challenges.

This review examines the potential of anaerobic membrane bioreactor (AnMBR) to serve as the core technology for simultaneous recovery of clean water, energy, and nutrient from wastewater. The potential is significant as AnMBR treatment can remove a board range of trace organic contaminants relevant to water reuse, convert organics in wastewater to biogas for subsequent energy production, and liberate nutrients to soluble forms (e.g. ammonia and phosphorus) for subsequent recovery for fertilizer production. Yet, there remain several significant challenges to the further development of AnMBR. These challenges evolve around the dilute nature of municipal wastewater, which entails the need for pre-concentrating wastewater prior to AnMBR, and hence, issues related to salinity build-up, accumulation of substances, membrane fouling, and membrane stability. Strategies to address these challenges are proposed and discussed. A road map for further research is also provided to guide future AnMBR development toward resource recovery.

Salinity build-up in osmotic membrane bioreactors: Causes, impacts, and potential cures.

Osmotic membrane bioreactor (OMBR), which integrates forward osmosis (FO) with biological treatment, has been developed to advance wastewater treatment and reuse. OMBR is superior to conventional MBR, particularly in terms of higher effluent quality, lower membrane fouling propensity, and higher membrane fouling reversibility. Nevertheless, advancement and future deployment of OMBR are hindered by salinity build-up in the bioreactor (e.g., up to 50 mS/cm indicated by the mixed liquor conductivity), due to high salt rejection of the FO membrane and reverse diffusion of the draw solution. This review comprehensively elucidates the relative significance of these two mechanisms towards salinity build-up and its associated effects in OMBR operation. Recently proposed strategies to mitigate salinity build-up in OMBR are evaluated and compared to highlight their potential in practical applications. In addition, the complementarity of system optimization and modification to effectively manage salinity build-up are recommended for sustainable OMBR development.

An anaerobic membrane bioreactor - membrane distillation hybrid system for energy recovery and water reuse: Removal performance of organic carbon, nutrients, and trace organic contaminants.

In this study, a direct contact membrane distillation (MD) unit was integrated with an anaerobic membrane bioreactor (AnMBR) to simultaneously recover energy and produce high quality water for reuse from wastewater. Results show that AnMBR could produce 0.3-0.5L/g CODadded biogas with a stable methane content of approximately 65%. By integrating MD with AnMBR, bulk organic matter and phosphate were almost completely removed. The removal of the 26 selected trace organic contaminants by AnMBR was compound specific, but the MD process could complement AnMBR removal, leading to an overall efficiency from 76% to complete removal by the integrated system. The results also show that, due to complete retention, organic matter (such as humic-like and protein-like substances) and inorganic salts accumulated in the MD feed solution and therefore resulted in significant fouling of the MD unit. As a result, the water flux of the MD process decreased continuously. Nevertheless, membrane pore wetting was not observed throughout the operation.

Effects of sulphur on the performance of an anaerobic membrane bioreactor: Biological stability, trace organic contaminant removal, and membrane fouling.

This study investigated the impact of sulphur content on the performance of an anaerobic membrane bioreactor (AnMBR) with an emphasis on the biological stability, contaminant removal, and membrane fouling. Removal of 38 trace organic contaminants (TrOCs) that are ubiquitously present in municipal wastewater by AnMBR was evaluated. Results show that basic biological performance of AnMBR regarding biomass growth and the removal of chemical oxygen demand (COD) was not affected by sulphur addition when the influent COD/SO42- ratio was maintained higher than 10. Nevertheless, the content of hydrogen sulphate in the produced biogas increased significantly and membrane fouling was exacerbated with sulphur addition. Moreover, the increase in sulphur content considerably affected the removal of some hydrophilic TrOCs and their residuals in the sludge phase during AnMBR operation. By contrast, no significant impact on the removal of hydrophobic TrOCs was noted with sulphur addition to AnMBR.

Biomimetic aquaporin membranes for osmotic membrane bioreactors: Membrane performance and contaminant removal.

In this study, we investigated the performance of an osmotic membrane bioreactor (OMBR) enabled by a novel biomimetic aquaporin forward osmosis (FO) membrane. Membrane performance and removal of 30 trace organic contaminants (TrOCs) were examined. Results show that the aquaporin FO membrane had better transport properties in comparison with conventional cellulose triacetate and polyamide thin-film composite FO membranes. In particular, the aquaporin FO membrane exhibited much lower salt permeability and thus smaller reverse salt flux, resulting in a less severe salinity build-up in the bioreactor during OMBR operation. During OMBR operation, the aquaporin FO membrane well complemented biological treatment for stable and excellent contaminant removal. All 30 TrOCs selected here were removed by over 85% regardless of their diverse properties. Such high and stable contaminant removal over OMBR operation also indicates the stability and compatibility of the aquaporin FO membrane in combination with activated sludge treatment.

Effects of salinity build-up on the performance of an anaerobic membrane bioreactor regarding basic water quality parameters and removal of trace organic contaminants.

The effects of elevated inorganic salt concentration on anaerobic membrane bioreactor (AnMBR) treatment regarding basic biological performance and trace organic contaminant (TrOC) removal were investigated. A set of 33 TrOCs were selected to represent pharmaceuticals, steroids, and pesticides in municipal wastewater. Results show potential adverse effects of increase in the bioreactor salinity to 15g/L (as NaCl) on the performance of AnMBR with respect to chemical oxygen demand removal, biogas production, and the removal of most hydrophilic TrOCs. Furthermore, a decrease in biomass production was observed as salinity in the bioreactor increased. The removal of most hydrophobic TrOCs was high and was not significantly affected by salinity build-up in the bioreactor. The accumulation of a few persistent TrOCs in the sludge phase was observed, but such accumulation did not vary significantly as salinity in the bioreactor increased.