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Chen Chen, Brittney N Nguyen, Gabriel Mitchell, Shally R Margolis, Darren Ma, Daniel A Portnoy
Listeriolysin O (LLO) is a cholesterol-dependent cytolysin that mediates escape of Listeria monocytogenes from a phagosome, enabling growth of the bacteria in the host cell cytosol. LLO contains a PEST-like sequence that prevents it from killing infected cells, but the mechanism involved is unknown. We found that the LLO PEST-like sequence was necessary to mediate removal of LLO from the interior face of the plasma membrane, where it coalesces into discrete puncta. LLO interacts with Ap2a2, an adaptor protein involved in endocytosis, via its PEST-like sequence, and Ap2a2-dependent endocytosis is required to prevent LLO-induced cytotoxicity...
June 13, 2018: Cell Host & Microbe
Zhimin Duan, Qing Chen, Leilei Du, Jianbo Tong, Song Xu, Rong Zeng, Yuting Ma, Xu Chen, Min Li
Autophagy machinery has roles in the defense against microorganisms such as Candida albicans . Lipidated LC3, the marker protein of autophagy, participates in the elimination of C. albicans by forming a single-membrane phagosome; this process is called LC3-associated phagocytosis (LAP). However, the influence of C. albicans on autophagic flux is not clear. In this study, we found that C. albicans inhibited LC3 turnover in macrophages. After the phagocytosis of C. albicans in macrophages, we observed fewer acridine orange-positive vacuoles and RFP-GFP-LC3 puncta without colocalization with phagocytized C...
2018: Oxidative Medicine and Cellular Longevity
Martina Nagel, Rudolf Winklbauer
The leading edge mesendoderm (LEM) of the Xenopus gastrula moves as an aggregate by collective migration. However, LEM cells on fibronectin in vitro show contact inhibition of locomotion by quickly retracting lamellipodia upon mutual contact. We found that a fibronectin-integrin-syndecan module acts between p21-activated kinase-1 upstream and ephrinB1 downstream to promote the contact-induced collapse of lamellipodia. To function in this module, fibronectin has to be present as puncta on the surface of LEM cells...
June 8, 2018: Development
Raksha Singh, Seonghee Lee, Laura Ortega, Elison Blancaflor, Vemanna S Ramu, Muthappa Senthil-Kumar, Clemencia M Rojas, Kirankumar S Mysore
Plants are naturally resistant to most pathogens through a broad and durable defense response called nonhost disease resistance. Nonhost disease resistance is a complex process that includes preformed physical and chemical barriers and induced responses. In spite of its importance, many components of nonhost disease resistance remain to be identified and characterized. Using virus-induced gene silencing in Nicotiana benthamiana, we discovered a novel gene that we named NbNHR2 (N. benthamiana nonhost resistance 2)...
June 7, 2018: Molecular Plant-microbe Interactions: MPMI
Gherardo Varando, Ruth Benavides-Piccione, Alberto Muñoz, Asta Kastanauskaite, Concha Bielza, Pedro Larrañaga, Javier DeFelipe
The development of 3D visualization and reconstruction methods to analyse microscopic structures at different levels of resolutions is of great importance to define brain microorganization and connectivity. MultiMap is a new tool that allows the visualization, 3D segmentation and quantification of fluorescent structures selectively in the neuropil from large stacks of confocal microscopy images. The major contribution of this tool is the posibility to easily navigate and create regions of interest of any shape and size within a large brain area that will be automatically 3D segmented and quantified to determine the density of puncta in the neuropil...
2018: Frontiers in Neuroanatomy
John P Wise, Charles G Price, Joseph A Amaro, Jason R Cannon
The motor features of Parkinson's disease (PD) primarily result from a lesion to the nigrostriatal dopamine system. Numerous non-motor symptoms occur in PD, many of which are postulated to stem from pathology outside of the nigrostriatal dopamine system. Perturbations to protein trafficking, disruption of mitochondrial integrity, and impaired autophagy have repeatedly been implicated in dopaminergic neuron cell death. Previously, we demonstrated that multiple markers of autophagy are disrupted in a rotenone model of PD, with alterations occurring prior to an overt lesion to the nigrostriatal dopamine system...
2018: Frontiers in Neuroscience
David Seung, Tina B Schreier, Léo Bürgy, Simona Eicke, Samuel C Zeeman
The mechanism of starch granule initiation in chloroplasts is not fully understood. Here we aimed to build on our recent discovery that PROTEIN TARGETING TO STARCH (PTST) family members, PTST2 and PTST3, are key players in starch granule initiation, by identifying and characterising additional proteins involved in the process in Arabidopsis chloroplasts. Using immunoprecipitation and mass spectrometry, we demonstrate that PTST2 interacts with two plastidial coiled-coil proteins. Surprisingly, one of the proteins is the thylakoid-associated MAR-BINDING FILAMENT-LIKE PROTEIN (MFP1), which was proposed to bind plastid nucleoids...
June 4, 2018: Plant Cell
Hua Yi, Kun Wang, Biaoyan Du, Lina He, Hiuting Ho, Maosong Qiu, Yidan Zou, Qiao Li, Junfeng Jin, Yujuan Zhan, Zhongxiang Zhao, Xiaodong Liu
Aleuritolic acid (AA) is a triterpene that is isolated from the root of Croton crassifolius Geisel. In the present study, the cytotoxic effects of AA on hepatocellular carcinoma cells were evaluated. AA exerted dose- and time-dependent cytotoxicity by inducing mitochondria-dependent apoptosis in the hepatocellular carcinoma cell line, HepG2. Meanwhile, treatment with AA also caused dysregulation of autophagy, as evidenced by enhanced conversion of LC3-I to LC3-II, p62 accumulation, and co-localization of GFP and mCherry-tagged LC3 puncta...
June 2, 2018: Molecules: a Journal of Synthetic Chemistry and Natural Product Chemistry
Marcus M Nalaskowski, Patrick Ehm, Christoph Rehbach, Nina Nelson, Maike Täger, Kathrin Modest, Manfred Jücker
The inositol 5-phosphatase SHIP1 acts as negative regulator of intracellular signaling in myeloid cells and is a tumor suppressor in myeloid leukemogenesis. After relocalization from the cytoplasm to the plasma membrane SHIP1 terminates PI3-kinase mediated signaling processes. Furthermore, SHIP1 is also found in distinct puncta in the cell nucleus and nuclear SHIP1 has a pro-proliferative function. Here we report the identification of five nuclear export signals (NESs) which regulate together with the two known nuclear localization signals (NLSs) the nucleocytoplasmic shuttling of SHIP1...
May 28, 2018: Cellular Signalling
Li Hua, Guirong Zhu, Jianguo Wei
Non-small cell lung cancer (NSCLC) is a major type of lung cancer. Drug resistance is a enormous obstacle for cancer treatment. Copious microRNAs (miRNAs) have been demonstrated to be implicated in drug resistance in NSCLC. In the present study, RT-qPCR assay revealed that microRNA-1 (miR-1) expression was downregulated in DDP resistant NSCLC tissues and cells. Western blot assay presented a remarkable increase of LC3B-II/LC3B-I ratio and a notable decline of p62 level in DDP resistant NSCLC cells, while these effects were weakened by miR-1...
May 30, 2018: Cell Biology International
Mohammed M Nooh, Salvatore Mancarella, Suleiman W Bahouth
The β1 -adrenergic receptor (β1 -AR) is a major cardiac G protein-coupled receptor, which mediates cardiac actions of catecholamines and is involved in genesis and treatment of numerous cardiovascular disorders. In mammalian cells, catecholamines induce the internalization of the β1 -AR into endosomes and their removal promotes the recycling of endosomal β1 -AR back to the plasma membrane, but whether these redistributive processes occur in terminally differentiated cells is unknown. Compartmentalization of the β1 -AR in response to β-agonists and antagonists was determined by confocal microscopy in primary adult rat ventricular myocytes (ARVM), which are terminally differentiated myocytes with unique structures such as t-tubules and contractile sarcomeres...
May 30, 2018: Molecular Pharmacology
Tae Woo Kim, Seon Young Lee, Mia Kim, Chunhoo Cheon, Bo-Hyoung Jang, Yong Cheol Shin, Seong-Gyu Ko
Danggui-Sayuk-Ga-Osuyu-Saenggang-Tang (DSGOST in Korean, Danggui-Sini-Jia-Wuzhuyu-Shengian-Tang in Chinese, and Tokishigyakukagoshuyushokyoto (TJ-38) in Japanese), a well-known traditional Korean/Chinese/Japanese medicine, has long been used to treat vascular diseases such as Raynaud's phenomenon (RP). However, anticancer effect of DSGOST remains elusive. In this study, we checked if DSGOST has an anticancer effect against gastric cancer cells, and investigated the mechanisms underlying DSGOST resistance. Moreover, DSGOST regulates chemoresistance in cisplatin-treated gastric cancer cells...
May 29, 2018: Cell Death & Disease
Okuno Ryoko, Yuko Ito, Nabil Eid, Yoshinori Otsuki, Yoichi Kondo, Koichi Ueda
Keloid is a fibro-proliferative skin disorder with tumor-like behavior and dependence on anaerobic glycolysis (the Warburg effect), but its exact pathogenesis is unknown. Although autophagy is widely accepted as a lysosomal pathway for cell survival and cellular homeostasis (specifically upon exposure to stressors such as hypoxia), very few studies have investigated the involvement of autophagy and related glycolytic effectors in keloidogenesis. Here the authors examined the expression and cellular localization of autophagy proteins (LC3, pan-cathepsin), glycolytic markers (LDH, MCT1, MCT4) and the transcription factor HIF isoforms in human keloid samples using immunohistochemical analysis and double-labeling immunofluorescence methods...
May 29, 2018: Histology and Histopathology
Yabin Hu, Yan Zhou, Gaoxiang Yang, Yalin Wang, Zhongnan Zheng, Juntao Li, Yuting Yan, Wei Wu
Sulforaphane-N-acetyl-cysteine (SFN-NAC) is a potential drug to inhibit human non-small cell lung cancer (NSCLC), but the underlying mechanisms are elusive. Here, we uncovered that SFN-NAC induced apoptosis via flow cytometer assay and transmission electron microscopy. Further, SFN-NAC increased LC3 II/LC3 I and the number of LC3 punctas, but Western blot showed that SFN-NAC inhibited cell autophagy in response to a co-treatment of Bafilomycin A1 and SFN-NAC. Furthermore, immunofluorescence staining and Western blot showed that SFN-NAC triggered microtubule disruption causing apoptosis via downregulating α-tubulin and phosphorylated ERK1/2-mediated Stathmin-1...
May 25, 2018: Cancer Letters
Hiroshi Nagahama, Masahito Nakazaki, Masanori Sasaki, Yuko Kataoka-Sasaki, Takahiro Namioka, Ai Namioka, Shinichi Oka, Rie Onodera, Junpei Suzuki, Yuichi Sasaki, Jeffery D Kocsis, Osamu Honmou
Systemic administration of mesenchymal stem cells (MSCs) following cerebral infarction exerts functional improvements. Previous research has suggested potential therapeutic mechanisms that promote neuroprotection and synaptogenesis. These include secretion of neurotrophic factors, remodeling of neural circuits, restoration of the blood brain barrier, reduction of inflammatory infiltration and demyelination, and elevation of trophic factors. In addition to these mechanisms, we hypothesized that restored interhemispheric bilateral motor cortex connectivity might be an additional mechanism of functional recovery...
May 23, 2018: Brain Research
Hollis McClory, Xiaolong Wang, Ellen Sapp, Leah W Gatune, Maria Iuliano, Chiu-Yi Wu, Gina Nathwani, Kimberly B Kegel-Gleason, Marian DiFiglia, Xueyi Li
Human huntingtin (Htt) contains 3144 amino acids and has an expanded polyglutamine region near the NH2 -terminus in patients with Huntington's disease. While numerous binding partners have been identified to NH2 -terminal Htt, fewer proteins are known to interact with C-terminal domains of Htt. Here we report that kalirin, a Rac1 activator, is a binding partner to C-terminal Htt. Kalirin and Htt co-precipitated from mouse brain endosomes and co-localized at puncta in NRK and immortalized striatal cells and primary cortical neurons...
May 22, 2018: Scientific Reports
Heather A Nelson, Colin A Leech, Richard F Kopp, Michael W Roe
Store-operated calcium entry (SOCE), a fundamentally important homeostatic and Ca2+ signaling pathway in many types of cells, is activated by the direct interaction of stromal interaction molecule 1 (STIM1), an endoplasmic reticulum (ER) Ca2+ -binding protein, with Ca2+ -selective Orai1 channels localized in the plasma membrane. While much is known about the regulation of SOCE by STIM1, the role of stromal interaction molecule 2 (STIM2) in SOCE remains incompletely understood. Here, using clustered regularly interspaced short palindromic repeats -CRISPR associated protein 9 (CRISPR-Cas9) genomic editing and molecular imaging, we investigated the function of STIM2 in NIH 3T3 fibroblast and αT3 cell SOCE...
May 19, 2018: International Journal of Molecular Sciences
Tian Zhang, Junkai Yan, Nan Wang, Lina Dai, Ying Wang, Wei Cai
BACKGROUND: The objectives of this study were to address the role of autophagy in the pathogenesis of parenteral nutrition (PN)-associated liver disease (PNALD) and its possible mechanism in vivo. METHODS: Five-week-old male Sprague Dawley rats were fed Shoobree chow (Xietong Organism, Jiangsu, China) and administered intravenous 0.9% saline (sham group), PN (PN group), PN plus rapamycin (1 mg/kg; PN + Rapa group), or rapamycin (Rapa group) for 7 days. Before and after study, body weight, biochemical indicators, hepatic histology, level of autophagy, hepatocyte apoptosis, reactive oxygen species (ROS), and endoplasmic reticulum (ER) stress indicators including binding immunoglobulin protein (BIP), spliced X-box-binding protein-1 (sXBP1), and CCAAT-enhancer-binding protein homologous protein (CHOP) were measured...
May 15, 2018: JPEN. Journal of Parenteral and Enteral Nutrition
Kui Xu, Xiu-Fen Liu, Zhi-Qiang Ke, Qing Yao, Shuang Guo, Chao Liu
BACKGROUND/AIMS: Diabetic cardiomyopathy is associated with increased apoptosis and suppressed autophagy in cardiac cells. The polyphenol resveratrol has shown beneficial effects in various cardiovascular diseases. This study investigated if resveratrol protected cardiac cells by modulating apoptosis and autophagy in the context of diabetes. METHODS: H9c2 cardiac myoblast cells were exposed to high glucose combined with palmitate. Autophagy was evaluated by estimating LC3-II/I ratio, P62 protein levels, and LC3 fluorescent puncta...
April 28, 2018: Cellular Physiology and Biochemistry
Shu-Yu Cheng, Nan-Fu Chen, Hsiao-Mei Kuo, San-Nan Yang, Chun-Sung Sung, Ping-Jyun Sung, Zhi-Hong Wen, Wu-Fu Chen
Prodigiosin, a secondary metabolite isolated from marine Vibrio sp., has antimicrobial and anticancer properties. This study investigated the cell death mechanism of prodigiosin in glioblastoma. Glioblastoma multiforme (GBM) is an aggressive primary cancer of the central nervous system. Despite treatment, or standard therapy, the median survival of glioblastoma patients is about 14.6 month. The results of the present study clearly showed that prodigiosin significantly reduced the cell viability and neurosphere formation ability of U87MG and GBM8401 human glioblastoma cell lines...
May 2, 2018: Apoptosis: An International Journal on Programmed Cell Death
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