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Nocodazole AND beta cells

Tao Wang, Ruping Wang, Rachel A Cleary, Olivia J Gannon, Dale D Tang
β-Catenin is a key component that connects transmembrane cadherin with the actin cytoskeleton at the cell-cell interface. However, the role of the β-catenin/cadherin interaction in smooth muscle has not been well characterized. Here stimulation with acetylcholine promoted the recruitment of β-catenin to N-cadherin in smooth muscle cells/tissues. Knockdown of β-catenin by lentivirus-mediated shRNA attenuated smooth muscle contraction. Nevertheless, myosin light chain phosphorylation at Ser-19 and actin polymerization in response to contractile activation were not reduced by β-catenin knockdown...
April 3, 2015: Journal of Biological Chemistry
Damien Stark, Joel L N Barratt, Tamalee Roberts, Deborah Marriott, John T Harkness, John Ellis
Recently, Dientamoeba fragilis has emerged as a significant and common enteropathogen. The majority of patients with dientamoebiasis present with gastrointestinal complaints and chronic symptoms are common. Numerous studies have successfully demonstrated parasite clearance, coupled with complete resolution of clinical symptoms following treatment with various antiparasitic compounds. Despite this, there is very little in vitro susceptibility data available for the organism. Benzimidazoles are a class of antiparasitic drugs that are commonly used for the treatment of protozoan and helminthic infections...
2014: Parasite: Journal de la Société Française de Parasitologie
R Suárez-Sánchez, A Aguilar, K M Wagstaff, G Velez, P M Azuara-Medina, P Gomez, A Vásquez-Limeta, O Hernández-Hernández, K G Lieu, D A Jans, B Cisneros
Even though the Duchenne muscular dystrophy (DMD) gene product Dystrophin Dp71d is involved in various key cellular processes through its role as a scaffold for structural and signalling proteins at the plasma membrane as well as the nuclear envelope, its subcellular trafficking is poorly understood. Here we map the nuclear import and export signals of Dp71d by truncation and point mutant analysis, showing for the first time that Dp71d shuttles between the nucleus and cytoplasm mediated by the conventional nuclear transporters, importin (IMP) α/β and the exportin CRM1...
May 2014: Biochimica et Biophysica Acta
Jiong Li, Xiaohong Chen, Xiangming Ding, Yingduan Cheng, Bin Zhao, Zhi-Chun Lai, Khalid Al Hezaimi, Razqallah Hakem, Kun-Liang Guan, Cun-Yu Wang
Abnormal activation of Wnt/β-catenin-mediated transcription is associated with a variety of human cancers. Here, we report that LATS2 inhibits oncogenic Wnt/β-catenin-mediated transcription by disrupting the β-catenin/BCL9 interaction. LATS2 directly interacts with β-catenin and is present on Wnt target gene promoters. Mechanistically, LATS2 inhibits the interaction between BCL9 and β-catenin and subsequent recruitment of BCL9, independent of LATS2 kinase activity. LATS2 is downregulated and inversely correlated with the levels of Wnt target genes in human colorectal cancers...
December 26, 2013: Cell Reports
Yusuke Kumai, Raymond W M Kwong, Steve F Perry
In the current study, the role of cAMP in stimulating Na(+) uptake in larval zebrafish was investigated. Treating larvae at 4 days postfertilization (dpf) with 10 μM forskolin or 1 μM 8-bromo cAMP significantly increased Na(+) uptake by three-fold and twofold, respectively. The cAMP-dependent stimulation of Na(+) uptake was probably unrelated to protein trafficking via microtubules because pretreatment with 200 μM colchicine or 30 μM nocodazole did not attenuate the magnitude of the response. Na(+) uptake was stimulated markedly following acute (2 h) exposure to acidic water...
January 1, 2014: American Journal of Physiology. Regulatory, Integrative and Comparative Physiology
Stephen L Bird, Rebecca Heald, Karsten Weis
Accurate positioning of the mitotic spindle is critical to ensure proper distribution of chromosomes during cell division. The small GTPase Ran, which regulates a variety of processes throughout the cell cycle, including interphase nucleocytoplasmic transport and mitotic spindle assembly, was recently shown to also control spindle alignment. Ran is required for the correct cortical localization of LGN and nuclear-mitotic apparatus protein (NuMA), proteins that generate pulling forces on astral microtubules (MTs) through cytoplasmic dynein...
August 2013: Molecular Biology of the Cell
Yukiko Kitase, Charles F Shuler
During palatal fusion, the midline epithelial seam (MES) degrades to achieve mesenchymal confluence. Epithelial mesenchymal transition (EMT) is one mechanism which is active in MES degradation. TGF-β induces EMT in medial edge epithelium (MEE) by down-regulation of an epithelial marker, E-cadherin. Microtubule disassembly impaired palatal fusion leading to a multi-layered MES in the mid-region. In this study, we investigated the effect of microtubule disruption on the regulation of the E-cadherin/catenin adhesion complex...
2013: International Journal of Developmental Biology
Ru Ya, Stephen M Downs
The oocyte meiotic spindle is comprised of microtubules (MT) that bind chromatin and regulate both metaphase plate formation and karyokinesis during meiotic maturation; however, little information is known about their role in meiosis reinitiation. This study was conducted to determine if microtubule integrity is required for meiotic induction and to ascertain how it affects activation of AMP-activated protein kinase (AMPK), an important participant in the meiotic induction process. Treatment with microtubule-disrupting agents nocodazole and vinblastine suppressed meiotic resumption in a dose-dependent manner in both arrested cumulus cell-enclosed oocytes (CEO) stimulated with follicle-stimulating hormone (FSH) and arrested denuded oocytes (DO) stimulated with the AMPK activator, 5-aminoimidazole-4-carboxamide-1-beta-4-ribofuranoside (AICAR)...
February 2014: Zygote: the Biology of Gametes and Early Embryos
Jens B Bosse, Rudolf Bauerfeind, Leonhard Popilka, Lisa Marcinowski, Martina Taeglich, Christophe Jung, Hannah Striebinger, Jens von Einem, Ulrike Gaul, Paul Walther, Ulrich H Koszinowski, Zsolt Ruzsics
Fluorescent tagging of viral particles by genetic means enables the study of virus dynamics in living cells. However, the study of beta-herpesvirus entry and morphogenesis by this method is currently limited. This is due to the lack of replication competent, capsid-tagged fluorescent viruses. Here, we report on viable recombinant MCMVs carrying ectopic insertions of the small capsid protein (SCP) fused to fluorescent proteins (FPs). The FPs were inserted into an internal position which allowed the production of viable, fluorescently labeled cytomegaloviruses, which replicated with wild type kinetics in cell culture...
2012: PloS One
Ming Shao, Yushuang Lin, Zhongzhen Liu, Ying Zhang, Lifeng Wang, Changbin Liu, Hongwei Zhang
The formation of dorsal-ventral (D-V) axis is the earliest event that breaks the radial symmetry and determines the bilateral body plan of a vertebrate embryo, however, the maternal control of this process is not fully understood. Here, we discovered a new dorsalizing window of acute lithium treatment, which covers only less than 10 minutes after fertilization. Lithium treatment in this window was not able to reverse the ventralized phenotype in tokkeabi (tkk) mutant embryos, and its dorsalizing activity on wild-type embryos was inhibited by nocodazole co-treatment...
2012: PloS One
Yukiko Kitase, Charles F Shuler
BACKGROUND: Formation of the secondary palate is complex and disturbance during palatal fusion may result in cleft palate. The processes of adhesion, intercalation, and disappearance of medial edge epithelia (MEE) are characterized by morphological changes requiring dynamic cytoskeletal rearrangement. Microtubules are one of the cytoskeletal elements involved in maintenance of cell morphology. Microtubule-disrupting drugs have been reported to cause craniofacial malformations including cleft palate...
July 2012: Developmental Dynamics: An Official Publication of the American Association of Anatomists
Hualong Yan, Songcheng Zhu, Chenlin Song, Naifa Liu, Jiuhong Kang
Aberrant expression of mitotic checkpoint genes compromises mitotic checkpoint, leads to chromosome instability and tumorigenesis. However, the cell signals that control mitotic checkpoint gene expression have not been reported so far. In the present study we show that, in human breast cancer cells, chemical inhibition of Bone morphogenetic proteins (BMPs), but not Transforming Growth Factor-β (TGF-β), abrogates the mitotic arrest induced by nocodazole. Protein expression analysis reveals that inhibition of BMP signaling dramatically down regulates protein levels of mitotic checkpoint components BUB3, Hec1, TTK and MAD2, but inhibition of TGF-β has relatively minor effect on the expression of these proteins...
April 2012: Cellular Signalling
Fu-I Lu, Christine Thisse, Bernard Thisse
In vertebrates, the animal-vegetal axis is determined during oogenesis and at ovulation, the egg is radially symmetric. In anamniotes, following fertilization, a microtubule-dependent movement leads to the displacement of maternal dorsal determinants from the vegetal pole to the future dorsal side of the embryo, providing the initial breaking of radial symmetry [Weaver C, Kimelman D (2004) Development 131:3491-3499]. These dorsal determinants induce β-catenin nuclear translocation in dorsal cells of the blastula...
September 20, 2011: Proceedings of the National Academy of Sciences of the United States of America
Kaizheng Gong, Dongqi Xing, Peng Li, Robert H Hilgers, Fadi G Hage, Suzanne Oparil, Yiu-Fai Chen
Atrial natriuretic peptide (ANP) and TGF-β play counterregulatory roles in pulmonary vascular adaptation to chronic hypoxia. We have demonstrated that ANP-cyclic GMP (cGMP)-protein kinase G (PKG) signaling inhibits TGF-β signaling by blocking TGF-β-induced nuclear translocation of mothers against decapentaplegic homolog (Smad)3 in pulmonary artery smooth muscle cells (PASMC). The current study tested the novel hypothesis that activation of the ANP-cGMP-PKG pathway limits TGF-β-induced Smad3 nuclear translocation by enhancing Smad3 binding to cytosolic anchoring proteins in isolated pulmonary artery smooth muscle cells...
October 2011: Molecular Endocrinology
Masahiro Nagahama, Yukari Itohayashi, Hideki Hara, Masahiro Higashihara, Yusuke Fukatani, Teruhisa Takagishi, Masataka Oda, Keiko Kobayashi, Ichiro Nakagawa, Jun Sakurai
The epsilon-toxin of Clostridium perfringens forms a heptamer in the membranes of Madin-Darby canine kidney cells, leading to cell death. Here, we report that it caused the vacuolation of Madin-Darby canine kidney cells. The toxin induced vacuolation in a dose-dependent and time-dependent manner. The monomer of the toxin formed oligomers on lipid rafts in membranes of the cells. Methyl-β-cyclodextrin and poly(ethylene glycol) 4000 inhibited the vacuolation. Epsilon-toxin was internalized into the cells. Confocal microscopy revealed that the internalized toxin was transported from early endosomes (early endosome antigen 1 staining) to late endosomes and lysosomes (lysosomal-associated membrane protein 2 staining) and then distributed to the membranes of vacuoles...
September 2011: FEBS Journal
Nabanita Biswas, Shufeng Liu, Tapani Ronni, Steven E Aussenberg, Weiqun Liu, Takashi Fujita, Tianyi Wang
BACKGROUND: The innate immune responses to virus infection are initiated by either Toll-like receptors (TLR3/7/8/9) or cytoplasmic double-stranded RNA (dsRNA)-recognizing RNA helicases RIG-I and MDA5. To avoid causing injury to the host, these signaling pathways must be switched off in time by negative regulators. METHODOLOGY/PRINCIPAL FINDINGS: Through yeast-two hybrid screening, we found that an ubiquitin-like protein named protein linking integrin-associated protein to cytoskeleton 1(PLIC-1 or Ubiquilin 1) interacted with the Toll/interleukin-1 receptor (TIR) domain of TLR4...
2011: PloS One
Enrico Ragni, Julia Calderon, Umberto Fascio, Matthias Sipiczki, William A Fonzi, Laura Popolo
Cell wall biogenesis is a dynamic process relying on the coordinated activity of several extracellular enzymes. PHR1 is a pH-regulated gene of Candida albicans encoding a glycosylphosphatidylinositol-anchored β(1,3)-glucanosyltransferase of family GH72 which acts as a cell wall remodelling enzyme and is crucial for morphogenesis and virulence. In order to explore the function of Phr1p, we obtained a green fluorescent protein (GFP) fusion to determine its localization. During induction of vegetative growth, Phr1p-GFP was concentrated in the plasma membrane of the growing bud, in the mother-bud neck, and in the septum...
August 2011: Fungal Genetics and Biology: FG & B
K Buki, E Kirsten, P Bauer, C Vidair, A Kun, J Mendeleyev, E Kun
The GTPase activity of purified dimeric tubulin (alpha+beta) at 5 mu M was insensitive to methyl-3,5-diiodo-4-(4'-methoxyphenoxy) benzoate (DIME), in contrast to nocodazole which activated GTPase. Cellular motility of MDA-MB-231 (human mammary cancer) cells migrating through 12-mu m pores was inhibited by DIME similar to nocodazole in a drug concentration-and DIME structure-dependent manner. An increase of cytoplasmic ATPase activity of DIME-treated cells without a decrease in ATP contents of intact cells suggests that DIME may also influence additional as yet unidentified ATP-dependent system(s) probably also involved in cell motility...
December 1997: International Journal of Oncology
N S Petrov, O V Zhidkova, V V Zenin, Iu M Rozanov, B V Popov
Suppressor complex p130/E2f4 inhibits transcription of multiple genes proteins regulating cell cycle progression and induces cell cycle arrest at G0/G1 required for induction of cell differentiation in cells of many tissues in vivo and various cell lineages in vitro. We found here that, in mesenchymal stem cells, (MSC) activation of the Wnt/beta-catenin signal pathway induced by MSC coculture with the A-549 cell line or by growth in the medium containing Li+ ions, which resulted in the accumulation of active forms of the p130, E2f4 and beta-catenin, was not coupled with inhibition of cell cycle progression...
2011: Tsitologiia
Eric Bernard, Maxime Solignat, Bernard Gay, Nathalie Chazal, Stephen Higgs, Christian Devaux, Laurence Briant
BACKGROUND: The replicative cycle of chikungunya virus (CHIKV), an alphavirus that recently re-emerged in India and in Indian Ocean area, remains mostly unknown. The aim of the present study was to investigate the intracellular trafficking pathway(s) hijacked by CHIKV to enter mammalian cells. METHODOLOGY/PRINCIPAL FINDINGS: Entry pathways were investigated using a variety of pharmacological inhibitors or overexpression of dominant negative forms of proteins perturbating cellular endocytosis...
2010: PloS One
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