The Hippo pathway plays a crucial role in cell proliferation and differentiation during tumorigenesis, tissue homeostasis and early embryogenesis. Scaffold proteins from the ezrin-radixin-moesin (ERM) family, including neurofibromin 2 (NF2; Merlin), regulate the Hippo pathway through cell polarity. However, the mechanisms underlying Hippo pathway regulation via cell polarity in establishing outer cells remain unclear. In this study, we generated artificial Nf2 mutants in the N-terminal FERM domain (L64P) and examined Hippo pathway activity by assessing the subcellular localization of YAP1 in early embryos expressing these mutant mRNAs. The L64P-Nf2 mutant inhibited NF2 localization around the cell membrane, resulting in YAP1 cytoplasmic translocation in the polar cells. L64P-Nf2 expression also disrupted the apical centralization of both large tumor suppressor 2 (LATS2) and ezrin in the polar cells. Furthermore, Lats2 mutants in the FERM binding domain (L83K) inhibited YAP1 nuclear translocation. These findings demonstrate that NF2 subcellular localization mediates cell polarity establishment involving ezrin centralization. This study provides previously unreported insights into how the orchestration of the cell-surface components, including NF2, LATS2 and ezrin, modulates the Hippo pathway during cell polarization.

Considerable attention has been paid in recent years to the recovery and effective utilization of organic matter in municipal wastewater for the establishment of a circular economy. Direct membrane filtration (DMF) of municipal wastewater using microfiltration (MF) or ultrafiltration (UF) membranes to retain and concentrate the organic matter in municipal wastewater could be a practical option for this purpose. However, severe membrane fouling and high concentrations of organic matter remaining in the DMF permeate are concerns to be addressed. Application of a simple pretreatment using fixed biofilms was investigated to address these issues. In this study, experiments were carried out at an existing municipal wastewater treatment plant. A moving bed biofilm reactor (MBBR) process operated under a very short HRT of 1 h and DO concentration of 0.5 mg/L selectively degraded low-molecular-weight dissolved organic matter in municipal wastewater without degradation of membrane-recoverable suspended and colloidal organic matter. Application of the pretreatment did not reduce the amount of organic carbon recovered by DMF using an MF membrane (approximately 70% of the influent COD being recovered), while it dramatically mitigated the membrane fouling probably due to the alteration of characteristics of dissolved organic matter in wastewater. The pretreatment also reduced the concentration of organic matter in the DMF permeate by 41%: COD concentration in the DMF permeate was as low as 40 mg/L. With the established MBBR pretreatment, performances of MF (0.1 mu m) and UF (molecular weight cut-off: 150,000) membranes for DMF were compared in parallel. It was found that the increase of the recoverable amount of organic matter by using UF was marginal (about 5%), whereas fouling in UF was much more severe than that in MF. The severe fouling in UF was caused by inorganic colloids such as FeS that could pass through MF membranes but be retained by UF membranes. Based on the results obtained in this study, it is concluded that MF is more suitable than UF for efficient DMF.

都市下水中の有機物と栄養塩(リン,窒素)は重要な資源である.都市下水から有機物を回収する方法の一つに下水直接膜ろ過(Direct Membrane Filtration, DMF)があり,流入下水中有機物の75-80%を安定して濃縮回収することができる.これまでに報告されているDMF検討例のほとんどで栄養塩の回収は行われず,処理水(膜透過水)に栄養塩が残存していた.本研究ではDMF前段においてリン吸着性能を付与した微生物担体を用いたリン吸着を,DMF後段においてイオン交換による膜透過水中アンモニアイオンの吸着を試みた.実下水を用いた実験で,リンに関しては75%,アンモニアイオンに関しては80%を回収できることが示された.アンモニアイオンの回収については,ナノろ過を導入して共存多価イオンの影響を排除することで,さらに回収率を高められることを確認できた.