'Dicumarol is a naturally occurring anticoagulant derived from coumarin that induces cytotoxicity and oxidative stress in human pancreatic cancer cells (Cullen, J. J., Hinkhouse, M. M., Grady, M., Gaut, A. W., Liu, J., Zhang, Y., Weydert, C. J. D., Domann, F. E., and Oberley, L. W. (2003) Cancer Res. 63, ... More
Profilin 1 is essential for retention and metabolism of mouse hematopoietic stem cells in bone marrow.
AuthorsZheng J, Lu Z, Kocabas F, Böttcher RT, Costell M, Kang X, Liu X, Deberardinis RJ, Wang Q, Chen GQ, Sadek H, Zhang CC,
Journal
PubMed ID24385538
'How stem cells interact with the microenvironment to regulate their cell fates and metabolism is largely unknown. Here we demonstrated that the deletion of the cytoskeleton-modulating protein profilin 1 (pfn1) in hematopoietic stem cell (HSCs) led to bone marrow failure, loss of quiescence, and mobilization and apoptosis of HSCs in ... More
KEAP1 is a redox sensitive target that arbitrates the opposing radiosensitive effects of parthenolide in normal and cancer cells.
'Elevated oxidative stress is observed more frequently in cancer cells than in normal cells. It is therefore expected that additional exposure to a low level of reactive oxygen species (ROS) will push cancer cells toward death, whereas normal cells might maintain redox homeostasis through adaptive antioxidant responses. We previously showed ... More
G-protein ligands inhibit in vitro reactions of vacuole inheritance.
AuthorsHaas A, Conradt B, Wickner W
JournalJ Cell Biol
PubMed ID8027189
'During budding in Saccharomyces cerevisiae, maternal vacuole material is delivered into the growing daughter cell via tubular or vesicular structures. One of the late steps in vacuole inheritance is the fusion in the bud of vesicles derived from the maternal vacuole. This process has been reconstituted in vitro and requires ... More
A trace amount of the human pro-apoptotic factor Bax induces bacterial death accompanied by damage of DNA.
AuthorsAsoh S, Nishimaki K, Nanbu-Wakao R, Ohta S
JournalJ Biol Chem
PubMed ID9556634
'An amount of human pro-apoptotic Bax as low as 0.01% of total protein was sufficient to cause cell death in Escherichia coli. The bacterial cell death was examined using a viable bacteria-specific fluorescence indicator system and loss of colony formation ability. Co-expression of anti-apoptotic Bcl-xL showed a modest inhibitory effect ... More
IGF-I regulates redox status in breast cancer cells by activating the amino acid transport molecule xC-.
AuthorsYang Y, Yee D,
Journal
PubMed ID24686172
'Insulin-like growth factors (IGF) stimulate cell growth in part by increasing amino acid uptake. xCT (SLC7A11) encodes the functional subunit of the cell surface transport system xC(-), which mediates cystine uptake, a pivotal step in glutathione synthesis and cellular redox control. In this study, we show that IGF-I regulates cystine ... More
Inducing the cell cycle arrest and apoptosis of oral KB carcinoma cells by hydroxychavicol: roles of glutathione and reactive oxygen species.
AuthorsChang MC, Uang BJ, Wu HL, Lee JJ, Hahn LJ, Jeng JH
JournalBr J Pharmacol
PubMed ID11834609
'Hydroxychavicol (HC; 10 - 50 microM), a betel leaf component, was found to suppress the 2% H(2)O(2)-induced lucigenin chemiluminescence for 53 - 75%. HC (0.02 - 2 microM) was also able to trap superoxide radicals generated by a xanthine/xanthine oxidase system with 38 - 94% of inhibition. Hydroxyl radicals-induced PUC18 ... More
Fluorescence microscopy methods for yeast.
AuthorsPringle JR, Preston RA, Adams AE, Stearns T, Drubin DG, Haarer BK, Jones EW
JournalMethods Cell Biol
PubMed ID2476649
Analysis of leukocyte function by flow cytometric techniques.
AuthorsStelzer GT
JournalCytometry Suppl
PubMed ID3076368
Methods for studying the yeast vacuole.
AuthorsRoberts CJ, Raymond CK, Yamashiro CT, Stevens TH
JournalMethods Enzymol
PubMed ID1706462
The VPS8 gene is required for localization and trafficking of the CPY sorting receptor in Saccharomyces cerevisiae.
AuthorsChen YJ, Stevens TH
JournalEur J Cell Biol
PubMed ID8864656
To better understand the process of protein sorting to the yeast vacuole, the VPS8 gene was identified and characterized. VPS8 encodes a membrane-associated hydrophilic protein of 135 kDa (Vps8p), which is required for the accurate sorting of the vacuolar hydrolase, carboxypeptidase Y (CPY). vps8 mutant cells missort and secrete CPY ... More
ATP-mediated glia signaling.
AuthorsCotrina ML, Lin JH, López-García JC, Naus CC, Nedergaard M
JournalJ Neurosci
PubMed ID10751435
Glia calcium signaling has recently been identified as a potent modulator of synaptic transmission. We show here that the spatial expansion of calcium waves is mediated by ATP and subsequent activation of purinergic receptors. Ectopic expression of gap junction proteins, connexins (Cxs), leads to an increase in both ATP release ... More
Mitochondrial permeability transition: a common pathway to necrosis and apoptosis.
AuthorsKim JS, He L, Lemasters JJ
JournalBiochem Biophys Res Commun
PubMed ID12729580
Opening of high conductance permeability transition pores in mitochondria initiates onset of the mitochondrial permeability transition (MPT). The MPT is a causative event, leading to necrosis and apoptosis in hepatocytes after oxidative stress, Ca(2+) toxicity, and ischemia/reperfusion. CsA blocks opening of permeability transition pores and protects cell death after these ... More
Increased membrane permeability of apoptotic thymocytes: a flow cytometric study.
We have recently developed a method for the separation and quantification of viable apoptotic cells without the need for permeabilisation or fixation of the cells. The method is based on the observation that apoptotic rat thymocytes fluoresce more brightly than normal cells after a brief incubation with the DNA binding ... More
Hydrogenosome metabolism is the key target for antiparasitic activity of resveratrol against Trichomonas vaginalis.
AuthorsMallo N, Lamas J, Leiro JM,
JournalAntimicrob Agents Chemother
PubMed ID23478970
Metronidazole (MDZ) and related 5-nitroimidazoles are the recommended drugs for treatment of trichomoniasis, a sexually transmitted disease caused by the protozoan parasite Trichomonas vaginalis. However, novel treatment options are needed, as recent reports have claimed resistance to these drugs in T. vaginalis isolates. In this study, we analyzed for the ... More
Advantages and limitations of commonly used methods to assay the molecular permeability of gap junctional intercellular communication.
AuthorsAbbaci M, Barberi-Heyob M, Blondel W, Guillemin F, Didelon J,
JournalBiotechniques
PubMed ID18611167
The role of gap junctional intercellular communication (GJIC) in regulation of normal growth and differentiation is becoming increasingly recognized as a major cellular function. GJIC consists of intercellular exchange of low molecular weight molecules, and is the only means for direct contact between cytoplasms of adjacent animal cells. Disturbances of ... More
Fluorescent fiber-optic calcium sensor for physiological measurements.
AuthorsShortreed M, Kopelman R, Kuhn M, Hoyland B
JournalAnal Chem
PubMed ID8651501
A new optical sensor based on covalent immobilization of a newly synthesized calcium-selective, long-wavelength, fluorescent indicator has been constructed, with a response dynamic range optimal for physiological measurements. Immobilization occurs via photoinitiated copolymerization of the indicator with acrylamide on the distal end of a silanized 125 micrograms diameter multimode optical ... More
Mdm31 and Mdm32 are inner membrane proteins required for maintenance of mitochondrial shape and stability of mitochondrial DNA nucleoids in yeast.
AuthorsDimmer KS, Jakobs S, Vogel F, Altmann K, Westermann B
JournalJ Cell Biol
PubMed ID15631992
The MDM31 and MDM32 genes are required for normal distribution and morphology of mitochondria in the yeast Saccharomyces cerevisiae. They encode two related proteins located in distinct protein complexes in the mitochondrial inner membrane. Cells lacking Mdm31 and Mdm32 harbor giant spherical mitochondria with highly aberrant internal structure. Mitochondrial DNA ... More
Characterization of VPS41, a gene required for vacuolar trafficking and high-affinity iron transport in yeast.
AuthorsRadisky DC, Snyder WB, Emr SD, Kaplan J
JournalProc Natl Acad Sci U S A
PubMed ID9159129
Mutations in the yeast gene VPS41 give rise to poor growth on low iron medium, severe alterations in vacuolar morphology, and cause the missorting of membranous and soluble vacuolar proteins. Our studies predict that VPS41 encodes a hydrophilic protein of 992 amino acids that contains no obvious signal sequence or ... More
In vitro reactions of vacuole inheritance in Saccharomyces cerevisiae.
AuthorsConradt B, Shaw J, Vida T, Emr S, Wickner W
JournalJ Cell Biol
PubMed ID1334958
Vacuole inheritance is temporally coordinated with the cell cycle and is restricted spatially to an axis between the maternal vacuole and the bud. The new bud vacuole is founded by a stream of vacuole-derived membranous vesicles and tubules which are transported from the mother cell into the bud to form ... More
Phosphorylation of p66Shc and forkhead proteins mediates Abeta toxicity.
Excessive accumulation of amyloid beta-peptide (Abeta) plays an early and critical role in synapse and neuronal loss in Alzheimer's Disease (AD). Increased oxidative stress is one of the mechanisms whereby Abeta induces neuronal death. Given the lessened susceptibility to oxidative stress exhibited by mice lacking p66Shc, we investigated the role ... More
Dicarboxy-dichlorofluorescein: a new fluorescent probe for measuring acidic intracellular pH.
AuthorsNedergaard M, Desai S, Pulsinelli W
JournalAnal Biochem
PubMed ID2372106
Derivatives of fluorescein sensitive to pH are extensively utilized for the determination of intracellular pH (pHi). Available dyes have pKa values of approximately 7.0, and are not well suited for measuring acidic pHi. We examined the fluorescein derivative, 5 (and 6)-carboxy-2',7'-dichlorofluorescein (CDCF) for its potential in the microspectrofluorometric measurement of ... More
A new class of mutants deficient in dodecamerization of aminopeptidase 1 and vacuolar transport.
AuthorsAndrei-Selmer C, Knuppel A, Satyanarayana C, Heese C, Schu PV
JournalJ Biol Chem
PubMed ID11152450
Vacuolar aminopeptidase 1 is transported to the vacuole by cytoplasmic double-membrane vesicles, the nonclassic Cvt pathway. The cytosolic protein dodecamerizes and is enclosed in a double-membrane vesicle, which is transported to and fuses with the vacuole releasing a single-membrane autophagic body into the vacuolar lumen. This is degraded and the ... More
Glucose deprivation-induced cytotoxicity and alterations in mitogen-activated protein kinase activation are mediated by oxidative stress in multidrug-resistant human breast carcinoma cells.
AuthorsLee YJ, Galoforo SS, Berns CM, Chen JC, Davis BH, Sim JE, Corry PM, Spitz DR
JournalJ Biol Chem
PubMed ID9478987
We previously observed that glucose deprivation induces cell death in multidrug-resistant human breast carcinoma cells (MCF-7/ADR). As a follow up we wished to test the hypothesis that metabolic oxidative stress was the causative process or at least the link between causative processes behind the cytotoxicity. In the studies described here, ... More
Ric1p and the Ypt6p GTPase function in a common pathway required for localization of trans-Golgi network membrane proteins.
AuthorsBensen ES, Yeung BG, Payne GS
JournalMol Biol Cell
PubMed ID11160819
In Saccharomyces cerevisiae, clathrin is necessary for localization of trans-Golgi network (TGN) membrane proteins, a process that involves cycling of TGN proteins between the TGN and endosomes. To characterize further TGN protein localization, we applied a screen for mutations that cause severe growth defects in combination with a temperature-sensitive clathrin ... More
Use of the anionic dye carboxy-2',7'-dichlorofluorescein for sensitive flow cytometric detection of multidrug resistance-associated protein activity.
AuthorsLaupeze B, Amiot L, Courtois A, Vernhet L, Drenou B, Fauchet R, Fardel O
JournalInt J Oncol
PubMed ID10427142
Multidrug resistance-associated protein (MRP) and P-glycoprotein are drug efflux pumps conferring multidrug resistance to tumor cells and sharing numerous substrates. In order to determine a flow cytometric assay allowing to analyse MRP activity in cancerous cells in a sensitive and specific manner, cellular accumulation and efflux of the anionic fluorescent ... More
The GTPase Ypt7p of Saccharomyces cerevisiae is required on both partner vacuoles for the homotypic fusion step of vacuole inheritance.
AuthorsHaas A, Scheglmann D, Lazar T, Gallwitz D, Wickner W
JournalEMBO J
PubMed ID7489715
In the budding yeast Saccharomyces cerevisiae, vacuoles are inherited by the projection of vesicles and tubules from the mother-cell vacuole into the growing daughter cell during the S phase. These vesicles then fuse and form the daughter-cell organelle. We have described previously in vitro reactions of the formation of vacuole-derived ... More
Yeast Vps45p is a Sec1p-like protein required for the consumption of vacuole-targeted, post-Golgi transport vesicles.
AuthorsPiper RC, Whitters EA, Stevens TH
JournalEur J Cell Biol
PubMed ID7720726
Over 45 VPS genes (vacuolar protein sorting) in Saccharomyces cerevisiae are necessary for the correct sorting and delivery of vacuolar hydrolases. Yeast strains carrying mutations in a subset of these VPS genes (class D vps mutants) are also defective in the segregation of vacuolar material into the developing daughter cell ... More
A new vital stain for visualizing vacuolar membrane dynamics and endocytosis in yeast.
AuthorsVida TA, Emr SD
JournalJ Cell Biol
PubMed ID7533169
We have used a lipophilic styryl dye, N-(3-triethylammoniumpropyl)-4- (p-diethylaminophenyl-hexatrienyl) pyridinium dibromide (FM 4-64), as a vital stain to follow bulk membrane-internalization and transport to the vacuole in yeast. After treatment for 60 min at 30 degrees C, FM 4-64 stained the vacuole membrane (ring staining pattern). FM 4-64 did not ... More
Characterization of calcium translocation across the plasma membrane of primary osteoblasts using a lipophilic calcium-sensitive fluorescent dye, calcium green C18.
AuthorsLloyd QP, Kuhn MA, Gay CV
JournalJ Biol Chem
PubMed ID7673232
The synthesis of Calcium Green C18, a lipophilic fluorescent calcium-sensitive dye, and its use as a monitor of Ca2+ efflux from cells is described. This indicator consists of a Calcium Green-1 molecule conjugated to a lipophilic 18-carbon alkyl chain which will intercalate into cell membranes. The Kd of the indicator ... More
Fluorescence microtopography of oxidative stress in lung ischemia-reperfusion.
Authorsal-Mehdi A, Shuman H, Fisher AB
JournalLab Invest
PubMed ID8176898
BACKGROUND: Generation of oxidizing radicals in lung ischemia-reperfusion (I/R) was evaluated with the oxidation-dependent fluorogen 2'-7' dichlorofluorescin diacetate (DCF-DA). EXPERIMENTAL DESIGN: Isolated, artificial media perfused, oxygen ventilated rat lungs were subjected to 1 hour of ischemia followed by 1 hour of reperfusion. DCF-DA (5 microM) present in 40 ml of ... More
Measurement of intracellular fluorescence of human monocytes relative to oxidative metabolism.
Human monocytes (MN) produce O2- and H2O2 when stimulated by agonists. Dichlorofluorescin diacetate (DCFH-DA) has been used as a substrate for measuring intracellular oxidant production in neutrophils. DCFH-DA is hydrolyzed by esterases to dichlorofluorescin (DCFH), which is trapped within the cell. This nonfluorescent molecule is then oxidized to fluorescent dichlorofluorescin ... More
Role of three rab5-like GTPases, Ypt51p, Ypt52p, and Ypt53p, in the endocytic and vacuolar protein sorting pathways of yeast.
AuthorsSinger-Krüger B, Stenmark H, Düsterhöft A, Philippsen P, Yoo JS, Gallwitz D, Zerial M
JournalJ Cell Biol
PubMed ID8163546
The small GTPase rab5 has been shown to represent a key regulator in the endocytic pathway of mammalian cells. Using a PCR approach to identify rab5 homologs in Saccharomyces cerevisiae, two genes encoding proteins with 54 and 52% identity to rab5, YPT51 and YPT53 have been identified. Sequencing of the ... More
Pharmacokinetics of 5 (and 6)-carboxy-2',7'-dichlorofluorescein and its diacetate promoiety in the liver.
Hepatic disposition of 5 (and 6)-carboxy-2',7'-dichlorofluorescein (CDF) and its diacetate promoiety (CDFDA) was studied in isolated perfused rat livers. Livers from Wistar wild-type and multidrug resistance-associated protein (Mrp)2-deficient (TR(-)) rats were perfused with CDF in the presence or absence of probenecid. Probenecid decreased the recovery of CDF in bile approximately ... More
Patch clamp studies on V-type ATPase of vacuolar membrane of haploid Saccharomyces cerevisiae. Preparation and utilization of a giant cell containing a giant vacuole.
AuthorsYabe I, Horiuchi K, Nakahara K, Hiyama T, Yamanaka T, Wang PC, Toda K, Hirata A, Ohsumi Y, Hirata R, Anraku Y, Kusaka I
JournalJ Biol Chem
PubMed ID10574964
A method for obtaining giant protoplasts of Escherichia coli (the spheroplast incubation (SI) method: Kuroda et al. (Kuroda, T., Okuda, N., Saitoh, N., Hiyama, T., Terasaki, Y., Anazawa, H., Hirata, A., Mogi, T., Kusaka, I., Tsuchiya, T., and Yabe, I. (1998) J. Biol. Chem. 273, 16897-16904) was adapted to haploid ... More
Molecular analysis of the yeast VPS3 gene and the role of its product in vacuolar protein sorting and vacuolar segregation during the cell cycle.
vps3 mutants of the yeast Saccharomyces cerevisiae are impaired in the sorting of newly synthesized soluble vacuolar proteins and in the acidification of the vacuole (Rothman, J. H., and T. H. Stevens. Cell. 47:1041-1051; Rothman, J. H., C. T. Yamashiro, C. K. Raymond, P. M. Kane, and T. H. Stevens. ... More
Novel PI(4)P 5-kinase homologue, Fab1p, essential for normal vacuole function and morphology in yeast.
AuthorsYamamoto A, DeWald DB, Boronenkov IV, Anderson RA, Emr SD, Koshland D
JournalMol Biol Cell
PubMed ID7663021
The FAB1 gene of budding yeast is predicted to encode a protein of 257 kDa that exhibits significant sequence homology to a human type II PI(4)P 5-kinase (PIP5K-II). The recently cloned human PIP5K-II specifically converts PI(4)P to PI(4,5)P2 (Boronenkov and Anderson, 1995). The region of highest similarity between Fab1p and ... More
Glutathione levels and sensitivity to apoptosis are regulated by changes in transaldolase expression.
AuthorsBanki K, Hutter E, Colombo E, Gonchoroff NJ, Perl A
JournalJ Biol Chem
PubMed ID8955144
Transaldolase (TAL) is a key enzyme of the reversible nonoxidative branch of the pentose phosphate pathway (PPP) that is responsible for the generation of NADPH to maintain glutathione at a reduced state (GSH) and, thus, to protect cellular integrity from reactive oxygen intermediates (ROIs). Formation of ROIs have been implicated ... More
Glutaminase and poly(ADP-ribose) polymerase inhibitors suppress pyrimidine synthesis and VHL-deficient renal cancers.
Authors
JournalJ Clin Invest
PubMed ID28346230
Epigenetic reprogramming governs EcSOD expression during human mammary epithelial cell differentiation, tumorigenesis and metastasis.
Authors
JournalOncogene
PubMed ID23318435
NADPH oxidase mediates radiation-induced oxidative stress in rat brain microvascular endothelial cells.
Authors
JournalFree Radic Biol Med
PubMed ID18640264
Neuronal XBP-1 Activates Intestinal Lysosomes to Improve Proteostasis in C. elegans.
Authors
JournalCurr Biol
PubMed ID31303493
The anti-tumour activity of DNA methylation inhibitor 5-aza-2'-deoxycytidine is enhanced by the common analgesic paracetamol through induction of oxidative stress.
Authors
JournalCancer Lett
PubMed ID33359448
Mitochondria are the major targets in albumin-induced apoptosis in proximal tubule cells.
Authors
JournalJ Am Soc Nephrol
PubMed ID17360944
Increased expression of NAPDH oxidase 4 in systemic sclerosis dermal fibroblasts: regulation by transforming growth factor β.
Authors
JournalArthritis Rheumatol
PubMed ID26096997
Distinct role of Hsp70 in Drosophila hemocytes during severe hypoxia.
Authors
JournalFree Radic Biol Med
PubMed ID21616137
ARID1B, a molecular suppressor of erythropoiesis, is essential for the prevention of Monge's disease.
Authors
JournalExp Mol Med
PubMed ID35672450
Stem cell differentiation with consistent lineage commitment induced by a flash of ultrafast-laser activation in vitro and in vivo.