Recognition of injured mitochondria for degradation by macroautophagy is essential for cellular health, but the mechanisms remain poorly understood. Cardiolipin is an inner mitochondrial membrane phospholipid. We found that rotenone, staurosporine, 6-hydroxydopamine and other pro-mitophagy stimuli caused externalization of cardiolipin to the mitochondrial surface in primary cortical neurons and SH-SY5Y cells. RNAi knockdown of cardiolipin synthase or of phospholipid scramblase-3, which transports cardiolipin to the outer mitochondrial membrane, decreased mitochondrial delivery to autophagosomes. Furthermore, we found that the autophagy protein microtubule-associated-protein-1-light chain-3 (LC3), which mediates both autophagosome formation and cargo recognition, contains cardiolipin-binding sites important for the engulfment of mitochondria by the autophagic system. Mutation of LC3 residues predicted as cardiolipin-interaction sites by computational modeling inhibited its participation in mitophagy. These data indicate that redistribution of cardiolipin serves as an “eat-me” signal for the elimination of damaged mitochondria from neuronal cells.
We report the identification and biotechnological utility of a plant gene encoding the tocopherol (vitamin E) biosynthetic enzyme 2-methyl-6-phytylbenzoquinol methyltransferase. This gene was identified by map-based cloning of the Arabidopsis mutation vitamin E pathway gene3-1 ( vte3-1 ), which causes increased accumulation of ␦ -tocopherol and decreased ␥ -tocopherol in the seed. Enzyme assays of recombinant protein supported the hypothesis that At-VTE3 encodes a 2-methyl-6-phytylbenzoquinol methyltransferase. Seed-specific expression of At-VTE3 in transgenic soybean reduced seed ␦ -tocopherol from 20 to 2%. These results confirm that At-VTE3 protein catalyzes the methylation of 2-methyl-6-phytylbenzoquinol in planta and show the utility of this gene in altering soybean tocopherol composition. When At-VTE3 was coexpressed with At-VTE4 ( ␥ -tocopherol methyltransferase) in soybean, the seed accumulated to Ͼ 95% ␣ -tocopherol, a dramatic change from the normal 10%, resulting in a greater than eightfold increase of ␣ -tocopherol and an up to fivefold increase in seed vitamin E activity. These findings demonstrate the utility of a gene identified in Arabidopsis to alter the tocopherol composition of commercial seed oils, a result with both nutritional and food quality implications.
Native cytochrome c (cyt c) has a compact tertiary structure with a hexacoordinated heme iron and functions in electron transport in mitochondria and apoptosis in the cytoplasm. However, the possibility that protein modifications confer additional functions to cyt c has not been explored.
Gliomas are the most common primary intracranial tumors. Understanding the molecular basis of gliomas' progression is required to develop more effective therapies. The Wnt/β-catenin signaling cascade is an important signal transduction pathway in human cancers. Although, overactivation of this pathway is a hallmark of several forms of cancer, little is known about its role in human gliomas. Here, we aimed to determine the clinical significance of Wnt/β-catenin pathway components in gliomas. Immunohistochemical staining was performed to detect the expression patterns of Wnt1, β-catenin and Cyclin D1 in the biopsies from 96 patients with primary gliomas. Kaplan-Meier survival and Cox regression analyses were performed to evaluate the prognosis of patients. Cytoplasmic staining pattern of Wnt1, membranous, cytoplasmic and nuclear accumulation of β-catenin, and nuclear localization of Cyclin D1 were demonstrated by immunohistochemical staining. The Wnt1 expression significantly correlated with the expression of Cyclin D1 (P < 0.0001). The ratio of tumors with a cytoplasmic-nuclear pattern or a cytoplasmic pattern of β-catenin was significantly higher in Wnt1-positive (P < 0.01) and Cyclin D1-positive (P < 0.01) tumors than in Wnt1-negative and Cyclin D1-negative tumors, respectively. The protein expression levels of Wnt1, β-catenin and Cyclin D1 were all positively correlated with the Karnofsky performance scale (KPS) score and World Health Organization (WHO) grades of patients with gliomas. Furthermore, Wnt1, cytoplasmic-nuclear β-catenin and Cyclin D1 status were all the independent prognostic factors for glioma patients (P = 0.01, 0.007 and 0.005, respectively). These results provide convincing evidence that the Wnt/β-catenin pathway correlated closely with the progression of gliomas and might be a novel prognostic marker for this neoplasm.
Among the distinct molecular signatures present in the mitochondrion is
the tetra-acylated anionic phospholipid cardiolipin, a lipid also present in
primordial, single-cell bacterial ancestors of mitochondria and multiple
bacterial species today. Cardiolipin is normally localized to the inner
mitochondrial membrane; however, when cardiolipin becomes externalized to the
surface of dysregulated mitochondria, it promotes inflammasome activation and
stimulates the elimination of damaged or nonfunctional mitochondria by
mitophagy. Given the immunogenicity of mitochondrial and bacterial membranes
that are released during sterile and pathogen-induced trauma, we hypothesized
that cardiolipins might function as “eat me” signals for
professional phagocytes. In experiments with macrophage cell lines and primary
macrophages, we found that membranes with mitochondrial or bacterial
cardiolipins on their surface were engulfed through phagocytosis, which depended
on the scavenger receptor CD36. Distinct from this process, the copresentation
of cardiolipin with the Toll-like receptor 4 (TLR4) agonist lipopolysaccharide
dampened TLR4-stimulated production of cytokines. These data suggest that
externalized, extracellular cardiolipins play a dual role in host-host and
host-pathogen interactions by promoting phagocytosis and attenuating
inflammatory immune responses.
Traction force microscopy provides a comprehensive description of the spatiotemporal dynamics of contractile activities and their regulation by guidance molecules in migrating neurons, as well as the underlying molecular mechanisms.
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