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Vahitha B Abdul-Salam, Giusy Russomanno, Chen Chien-Nien, Abdul S Mahomed, Luke A Yates, Martin R Wilkins, Lan Zhao, Magdalena Maria Gierula, Olivier Dubois, Ute Schaeper, Jens Endruschat, Beata Wojciak-Stothard
RATIONALE: Increased expression of chloride intracellular channel 4 (CLIC4) is a feature of endothelial dysfunction in pulmonary arterial hypertension but its role in disease pathology is not fully understood. OBJECTIVE: To identify CLIC4 effectors and evaluate strategies targeting CLIC4 signalling in pulmonary hypertension. METHODS AND RESULTS: Proteomic analysis of CLIC4-interacting proteins in human pulmonary artery endothelial cells (HPAECs) identified regulators of endosomal trafficking, including ADP Ribosylation Factor 6 (Arf6) GTPase activating proteins and clathrin, while CLIC4 overexpression affected protein regulators of vesicular trafficking, lysosomal function and inflammation...
October 15, 2018: Circulation Research
Alan Costello, Orla Coleman, Nga T Lao, Michael Henry, Paula Meleady, Niall Barron, Martin Clynes
miRNAs are potent molecular regulators of cellular behaviour. The manipulation of these small non-coding RNAs has been used to enhance industrially relevant phenotypes in Chinese Hamster Ovary (CHO) cells. We investigated the stable depletion of six miRNAs; miR-204-5p, 338-3p, 378-3p, 409-3p, 455-3p and 505-3p, robustly associated with cell growth rate from a previous profiling study. Inhibition of endogenous miR-378-3p function by miRNA-sponge-decoy improved peak cell density by 59%. Quantitative label free LC-MS/MS proteomic analysis of the fractionated cell cultures at day 4 and 8 of batch culture found 216 cytosolic and 114 membrane-associated proteins differentially expressed with stable miR-378-3p depletion...
October 31, 2018: Journal of Biotechnology
Elisabetta Argenzio, Jeffrey Klarenbeek, Katarzyna M Kedziora, Leila Nahidiazar, Tadamoto Isogai, Anastassis Perrakis, Kees Jalink, Wouter H Moolenaar, Metello Innocenti
Chloride intracellular channel 4 (CLIC4) is a cytosolic protein implicated in diverse actin-based processes, including integrin trafficking, cell adhesion and tubulogenesis. CLIC4 is rapidly recruited to the plasma membrane by RhoA-activating agonists and then partly co-localizes with β1 integrins. Agonist-induced CLIC4 translocation depends on actin polymerization and requires conserved residues that make up a putative binding groove. However, the mechanism and significance of CLIC4 trafficking have been elusive...
October 31, 2018: Journal of Biological Chemistry
Bipradeb Singha, Sandra L Harper, Aaron R Goldman, Benjamin G Bitler, Katherine M Aird, Mark E Borowsky, Mark G Cadungog, Qin Liu, Rugang Zhang, Stephanie Jean, Ronny Drapkin, David W Speicher
New plasma and tissue biomarkers of epithelial ovarian cancer (EOC) could improve early diagnosis and post-diagnosis clinical management. Here we investigated tissue staining and tissue secretion of CLIC1 and CLIC4 across EOC subtypes. CLIC1 and CLIC4 are two promising biomarkers we previously showed were elevated in EOC patient sera. Individually, CLIC1 or CLIC4 stained larger percentages of malignant tumors across all EOC subtypes compared with CA125, particularly early stage and mucinous tumors. CLIC4 also stained benign tumors but staining was limited to nuclei; whereas malignant tumors showed diffuse cellular staining of stromal and tumor cells...
October 3, 2018: Scientific Reports
Dong Guo, Wenting Xie, Pan Xiong, Huifang Li, Siqi Wang, Guimiao Chen, Yuehong Gao, Jiechao Zhou, Ye Zhang, Guojun Bu, Maoqiang Xue, Jie Zhang
Oxidative stress can cause apoptosis in neurons and may result in neurodegenerative diseases. However, the signaling mechanisms leading to oxidative stress-induced neuronal apoptosis are not fully understood. Oxidative stress stimulates aberrant activation of cyclin-dependent kinase 5 (CDK5), thought to promote neuronal apoptosis by phosphorylating many cell death-related substrates. Here, using protein pulldown methods, immunofluorescence experiments and in vitro kinase assays, we identified chloride intracellular channel 4 (CLIC4), the expression of which increases during neuronal apoptosis, as a CDK5 substrate...
September 20, 2018: Cell Death & Disease
Jun Liang, Aijo De Castro, Lizette Flores
In this protocol, a green fluorescence protein (GFP) fusion protein and 4',6-diamidino-2-phenylindole (DAPI) staining are used to track protein subcellular localization changes; in particular, a nuclear translocation under a heat stress condition. Proteins react correspondingly to external and internal signals. A common mechanism is to change its subcellular localization. This article describes a protocol to track protein localization that does not require an antibody, radioactive labeling, or a confocal microscope...
July 30, 2018: Journal of Visualized Experiments: JoVE
Jinsen Lu, Lele Wu, Xiaoke Wang, Jinhang Zhu, Juan Du, Bing Shen
Depletion of the mitochondrial membrane potential (MMP, ΔΨm) is considered the earliest event in the apoptotic cascade. It even occurs ahead of nuclear apoptotic characteristics, including chromatin condensation and DNA breakage. Once the MMP collapses, cell apoptosis will initiate irreversibly. A series of lipophilic cationic dyes can pass through the cell membrane and aggregate inside the matrix of mitochondrion, and serve as fluorescence marker to evaluate MMP change. As one of the six members of the Cl- intracellular channel (CLIC) family, CLIC4 participates in the cell apoptotic process mainly through the mitochondrial pathway...
July 17, 2018: Journal of Visualized Experiments: JoVE
Xiangyun Yin, Jixiu Zhao, Hong Jiang, Liangliang Li, Jian Jiang, Hongmin Xi, Xiangli Peng, Xiaohang Yin, Xiaotong Shi, Lulu Zhang
BACKGROUND: Premature birth is a significant health care burden. Xenon (Xe) is a general anesthetic with neuroprotective effects. OBJECTIVES: Here, we investigate the neuroprotective role of Xe in a lipopolysaccharide (LPS)- and hypoxia-ischemia (HI)-induced white matter damage (WMD) model. METHODS: Three-day-old Sprague-Dawley rats were randomly divided into a sham group (group A, n = 24), an LPS + HI group (group B, n = 24), and an LPS + HI + Xe group (group C, n = 72)...
2018: Neonatology
Gabrielė Stakaitytė, Nnenna Nwogu, Jonathan D Lippiat, G Eric Blair, Krzysztof Poterlowicz, James R Boyne, Andrew Macdonald, Jamel Mankouri, Adrian Whitehouse
Ion channels regulate many aspects of cell physiology, including cell proliferation, motility, and migration, and aberrant expression and activity of ion channels is associated with various stages of tumor development, with K+ and Cl- channels now being considered the most active during tumorigenesis. Accordingly, emerging in vitro and preclinical studies have revealed that pharmacological manipulation of ion channel activity offers protection against several cancers. Merkel cell polyomavirus (MCPyV) is a major cause of Merkel cell carcinoma (MCC), primarily because of the expression of two early regulatory proteins termed small and large tumor antigens (ST and LT, respectively)...
March 23, 2018: Journal of Biological Chemistry
Diane Hatziioanou, Georgios Barkas, Elena Critselis, Jerome Zoidakis, Hariklia Gakiopoulou, Maria-Eleni Androutsou, Garyfalia Drossopoulou, Aristidis Charonis, Demetrios V Vlahakos
BACKGROUND: Hypertensive nephropathy, a leading cause of declining kidney function, is a multifactorial process not well understood. In order to elucidate biological processes and identify novel macromolecular components crucially involved in the process of kidney damage, the application of system biology approaches, like proteomics, is required. METHODS: Proteomic studies were performed using the renal parenchyma of spontaneously hypertensive rats (SHR) and their normotensive Wistar Kyoto controls...
2018: Nephron
Qiu-Yun Yu, Xin-Feng Zhou, Qing Xia, Jia Shen, Jia Yan, Jiu-Ting Zhu, Xiang Li, Ming Shu
This study explored the effects involved in silencing CLIC4 on apoptosis and proliferation of mouse liver cancer Hca-F and Hca-P cells. A CLIC4-target small interfering RNA (siRNA) was designed to compound into two individual complementary oligonucleotide chains. A process of annealing and connection to a pSilencer vector was followed by transfection with Hca-F and Hca-P cells. Quantitative real-time polymerase chain reaction and Western blotting techniques were used to determine CLIC4 mRNA and protein expressions...
January 2018: Journal of Cellular Biochemistry
Raquel Domingo-Fernández, Rebecca C Coll, Jay Kearney, Samuel Breit, Luke A J O'Neill
The NLRP3 inflammasome is a multiprotein complex that regulates the activation of caspase-1 leading to the maturation of the proinflammatory cytokines IL-1β and IL-18 and promoting pyroptosis. Classically, the NLRP3 inflammasome in murine macrophages is activated by the recognition of pathogen-associated molecular patterns and by many structurally unrelated factors. Understanding the precise mechanism of NLRP3 activation by such a wide array of stimuli remains elusive, but several signaling events, including cytosolic efflux and influx of select ions, have been suggested...
July 21, 2017: Journal of Biological Chemistry
Yong Zhang, Chen Ma, Jie Zhao, Haiyan Xu, Qiangchuan Hou, Heping Zhang
The incidence of colon cancer has increased considerably and the intestinal microbiota participate in the development of colon cancer. We showed that the L. casei Zhang or vitamin K2 (Menaquinone-7) intervention significantly alleviated intestinal tumor burden in mice. This was associated with increased serum adiponectin levels in both treatments. But osteocalcin level was only increased by L. casei Zhang. Furthermore, the anti-carcinogenic actions of L. casei Zhang were mediated by hepatic Chloride channel-3(CLCN3)/Nuclear Factor Kappa B(NF-κB) and intestinal Claudin15/Chloride intracellular channel 4(CLIC4)/Transforming Growth Factor Beta(TGF-β) signaling, while the vitamin K2 effect involved a hepatic Vitamin D Receptor(VDR)-phosphorylated AMPK signaling pathway...
April 11, 2017: Oncotarget
Qiong Zou, Zhulin Yang, Daiqiang Li, Ziru Liu, Yuan Yuan
Pancreatic cancer is the fourth most common cause of cancer-related mortality. Novel molecular biomarkers need to be identified for personalized medicine and to improve survival. The aim of this study was to examine chloride intracellular channel 4 (CLIC4) and Indian Hedgehog (Ihh) expression in benign and malignant lesions of the pancreas and to examine the eventual association between CLIC4 and Ihh expression, with clinicopathological features and prognosis of pancreatic cancer. A retrospective study of specimens collected from January 2000 to December 2011 at the Department of Pathology of the Second and Third Xiangya Hospitals, Central South University was undertaken to explore this question...
December 2016: International Journal of Experimental Pathology
(no author information available yet)
[This corrects the article DOI: 10.1371/journal.pone.0161410.].
2016: PloS One
Mahtab Tavasoli, Abass Al-Momany, Xin Wang, Laiji Li, John C Edwards, Barbara J Ballermann
The chloride intracellular channel (CLIC) 5A is expressed at very high levels in renal glomeruli, in both endothelial cells (EC) and podocytes. CLIC5A stimulates Rac1- and phosphatidylinositol (4,5)-bisphosphate-dependent ERM (ezrin, radixin, moesin) activation. ERM proteins, in turn, function in lumen formation and in the development of actin-based cellular projections. In mice lacking CLIC5A, ERM phosphorylation is profoundly reduced in podocytes, but preserved in glomerular EC. Since glomerular EC also express CLIC4, we reasoned that, if CLIC4 activates ERM proteins like CLIC5A, then CLIC4 could compensate for the CLIC5A loss in glomerular EC...
November 1, 2016: American Journal of Physiology. Renal Physiology
Anjali Shukla, Yihan Yang, Sara Madanikia, Yan Ho, Mangmang Li, Vanesa Sanchez, Christophe Cataisson, Jing Huang, Stuart H Yuspa
CLIC4 (Chloride intracellular channel 4) belongs to a family of putative intracellular chloride channel proteins expressed ubiquitously in multiple tissues. CLIC4 is predominantly soluble and traffics between the cytoplasm and nucleus and participates in cell cycle control and differentiation. Transforming growth factor beta (TGF-β) elevates CLIC4, which enhances TGF-β signaling through CLIC4 mediated stabilization of phospho-Smad2/3. CLIC4 is essential for TGF-β induced conversion of fibroblasts to myofibroblasts and expression of matrix proteins, signaling via the p38MAPK pathway...
2016: PloS One
Devasena Ponnalagu, Shubha Gururaja Rao, Jason Farber, Wenyu Xin, Ahmed Tafsirul Hussain, Kajol Shah, Soichi Tanda, Mark A Berryman, John C Edwards, Harpreet Singh
Chloride intracellular channel (CLICs) proteins show 60-70% sequence identity to each other, and exclusively localize to the intracellular organelle membranes and cytosol. In support of our recent publication, "Molecular identity of cardiac mitochondrial chloride intracellular channel proteins" (Ponnalagu et al., 2016) [1], it was important to characterize the specificity of different CLIC paralogs/ortholog (CLIC1, CLIC4, CLIC5 and DmCLIC) antibodies used to decipher their localization in cardiac cells. In addition, localization of CLICs in the other organelles such as endoplasmic reticulum (ER) of cardiomyocytes was established...
June 2016: Data in Brief
Haowei Xue, Jinsen Lu, Renxiang Yuan, Jinli Liu, Yehai Liu, Kaile Wu, Jing Wu, Juan Du, Bing Shen
BACKGROUND: Human head and neck squamous carcinoma is the 6th most prevalent carcinoma worldwide. Although many novel therapies have been developed, the clinical treatment for patients remains non-ideal. Chloride intracellular channel 4 (CLIC4), one of the seven members of the CLIC family, is a newly found Cl(-) channel that participates in various biological processes, including cellular apoptosis and differentiation. Accumulating evidence has revealed the significant role of CLIC4 in regulating the apoptosis of different cancer cells...
2016: Cell & Bioscience
Szu-Yi Chou, Kuo-Shun Hsu, Wataru Otsu, Ya-Chu Hsu, Yun-Cin Luo, Celine Yeh, Syed S Shehab, Jie Chen, Vincent Shieh, Guo-an He, Michael B Marean, Diane Felsen, Aihao Ding, Dix P Poppas, Jen-Zen Chuang, Ching-Hwa Sung
Chloride intracellular channel 4 (CLIC4) is a mammalian homologue of EXC-4 whose mutation is associated with cystic excretory canals in nematodes. Here we show that CLIC4-null mouse embryos exhibit impaired renal tubulogenesis. In both developing and developed kidneys, CLIC4 is specifically enriched in the proximal tubule epithelial cells, in which CLIC4 is important for luminal delivery, microvillus morphogenesis, and endolysosomal biogenesis. Adult CLIC4-null proximal tubules display aberrant dilation. In MDCK 3D cultures, CLIC4 is expressed on early endosome, recycling endosome and apical transport carriers before reaching its steady-state apical membrane localization in mature lumen...
2016: Nature Communications
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