Search Results (4)

Ubiquitin-like modifier-activating enzyme 6 (UBA6) is a member of the E1 enzyme family, which initiates the ubiquitin–proteasome system (UPS). The UPS plays critical roles not only in protein degradation but also in various cellular functions, including neuronal signaling, myocardial remodeling, immune cell differentiation, and cancer development. However, the specific role of UBA6 in cellular functions is not fully elucidated in comparison with the roles of the UPS. It has been known that the E1 enzyme is associated with the motility of cancer cells. In this study, we verified the physiological roles of UBA6 in lung cancer cells through gene-silencing siRNA targeting UBA6 (siUBA6). The siUBA6 treatment attenuated the migration of H1975 cells, along with a decrease in lysosomal Ca²⁺ release. While autophagosomal proteins remained unchanged, lysosomal proteins, including TRPML1 and TPC2, were decreased in siUBA6-transfected cells. Moreover, siUBA6 induced the production of multivesicular bodies (MVBs), accompanied by an increase in MVB markers in siUBA6-transfected H1975 cells. Additionally, the expression of the exosomal marker CD63 and extracellular vesicles was increased by siUBA6 treatment. Our findings suggest that knock-down of UBA6 induces lysosomal TRPML1 depletion and inhibits endosomal trafficking to lysosome, and subsequently, leads to the accumulation of MVBs and enhanced exosomal secretion in lung cancer cells. Full article

Apoptosis-linked gene 2 (ALG-2, also known as PDCD6) is a member of the penta-EF-hand (PEF) family of Ca²⁺-binding proteins. The murine gene encoding ALG-2 was originally reported to be an essential gene for apoptosis. However, the role of ALG-2 in cell death pathways has remained elusive. In the present study, we found that cell death-inducing p53 target protein 1 (CDIP1), a pro-apoptotic protein, interacts with ALG-2 in a Ca²⁺-dependent manner. Co-immunoprecipitation analysis of GFP-fused CDIP1 (GFP-CDIP1) revealed that GFP-CDIP1 associates with tumor susceptibility gene 101 (TSG101), a known target of ALG-2 and a subunit of endosomal sorting complex required for transport-I (ESCRT-I). ESCRT-I is a heterotetrameric complex composed of TSG101, VPS28, VPS37 and MVB12/UBAP1. Of diverse ESCRT-I species originating from four VPS37 isoforms (A, B, C, and D), CDIP1 preferentially associates with ESCRT-I containing VPS37B or VPS37C in part through the adaptor function of ALG-2. Overexpression of GFP-CDIP1 in HEK293 cells caused caspase-3/7-mediated cell death. In addition, the cell death was enhanced by co-expression of ALG-2 and ESCRT-I, indicating that ALG-2 likely promotes CDIP1-induced cell death by promoting the association between CDIP1 and ESCRT-I. We also found that CDIP1 binds to vesicle-associated membrane protein-associated protein (VAP)A and VAPB through the two phenylalanines in an acidic tract (FFAT)-like motif in the C-terminal region of CDIP1, mutations of which resulted in reduction of CDIP1-induced cell death. Therefore, our findings suggest that different expression levels of ALG-2, ESCRT-I subunits, VAPA and VAPB may have an impact on sensitivity of anticancer drugs associated with CDIP1 expression. Full article

Although some studies have been conducted over the past few decades, the existence of mitochondria-rich cells (MRCs) in reptiles is still obscure. This is the first study to uncover the presence of MRCs in the small intestine of Chinese soft-shelled turtles. In this study, we investigated the ultrastructural characteristics of MRCs and the secretion of different ion transport proteins in the small intestine of Pelodiscus sinensis. Transmission electron microscopy revealed that the ultrastructural features of MRCs are clearly different from those of other cells. The cytoplasmic density of MRCs was higher than absorptive epithelial cells (AECs) and goblet cells (GCs). MRCs possessed abundant heterogeneous mitochondria and an extensive tubular system in the cytoplasm, however, the AECs and GCs completely lacked a tubular system. Statistical analysis showed that the diameter and quantification of mitochondria were highly significant in MRCs. Mitochondrial vacuolization and despoiled mitochondria were closely associated with autophagosomes in MRCs. The multivesicular bodies (MVBs) and the exosome secretion pathway were observed in MRCs. Immunohistochemical staining of ion transport proteins indicated positive immunoreactivity of Na⁺/K^+_ATPase (NKA) and Na⁺/K⁺/2Cl⁻ cotransporter (NKCC) at the basal region of the mucosal surface. Likewise, the immunofluorescence staining results showed a strong positive localization of NKA, NKCC, and carbonic anhydrase (CA) at the basal and apical region of the mucosal surface of small intestine. Our findings suggest that MRCs provide support and regulate cellular ions for intestinal homeostasis and provide energy for cellular quality control in intestine. Full article

Large-scale models are generally associated with large spatial modelling units, for example, counties or super grids (several to dozens of km²). Few applied urban models can achieve a large spatial coverage with irregular spatial units due to data availability and computation load. The framework of automatic identification and characterization of blocks developed by Liu and Long (2016) makes such an ideal model possible by establishing the existing urban blocks using road networks and points of interest for very large areas (e.g., a country or a continent). In this study, we develop a mega-vector-blocks cellular automata model (MVB-CA) to simulate urban expansion at the block level for 654 Chinese cities. The existing urban blocks in 2012 were used for initiating the MVB-CA and are generated using multi-levelled road networks and ubiquitous points of interest. We then simulate block-based urban expansion of all the cities from 2012 to 2017. The national spatial development strategies of China are discussed extensively by academia and policy makers, while the baseline scenario and other simulated urban expansion scenarios have been tested and compared horizontally. As one of the first block-based urban expansion models at a national scale, its academic contributions, practical applications, and potential biases are also discussed in this paper. The developed MVB-CA using general approaches is also applicable for other counties. Full article