Gastric Parietal Cell Physiology and Helicobacter pylori–Induced Disease

Yao, Xuebiao; Smolka, Adam J.

doi:10.1053/j.gastro.2019.02.036

Cited by 81 publications

(71 citation statements)

References 162 publications

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“…The excitement and challenge ahead are to delineate the molecular mechanism underlying context-dependent function of NDP52 in cell fate decision control. 47 Furthermore, another actin nucleation factor Arp2/3 that cooperates with N-WASP to promote actin polymerization may also be involved in the NDP52-mediated coordination between actin and microtubule networks. Future studies are needed to explore the potential role of Arp2/3 in this pathway.…”

Section: Discussionmentioning

confidence: 99%

NDP52 tunes cortical actin interaction with astral microtubules for accurate spindle orientation

Yang

Dong

et al. 2019

Cell Res

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Oriented cell divisions are controlled by a conserved molecular cascade involving Gαi, LGN, and NuMA. Here, we show that NDP52 regulates spindle orientation via remodeling the polar cortical actin cytoskeleton. siRNA-mediated NDP52 suppression surprisingly revealed a ring-like compact subcortical F-actin architecture surrounding the spindle in prophase/prometaphase cells, which resulted in severe defects of astral microtubule growth and an aberrant spindle orientation. Remarkably, NDP52 recruited the actin assembly factor N-WASP and regulated the dynamics of the subcortical F-actin ring in mitotic cells. Mechanistically, NDP52 was found to bind to phosphatidic acid-containing vesicles, which absorbed cytoplasmic N-WASP to regulate local filamentous actin growth at the polar cortex. Our TIRFM analyses revealed that NDP52-containing vesicles anchored N-WASP and shortened the length of actin filaments in vitro. Based on these results we propose that NDP52-containing vesicles regulate cortical actin dynamics through N-WASP to accomplish a spatiotemporal regulation between astral microtubules and the actin network for proper spindle orientation and precise chromosome segregation. In this way, intracellular vesicles cooperate with microtubules and actin filaments to regulate proper mitotic progression. Since NDP52 is absent from yeast, we reason that metazoans have evolved an elaborate spindle positioning machinery to ensure accurate chromosome segregation in mitosis.

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Section: Discussionmentioning

confidence: 99%

NDP52 tunes cortical actin interaction with astral microtubules for accurate spindle orientation

Yang

Dong

et al. 2019

Cell Res

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“…The members of the ezrin/radixin/moesin (ERM) family proteins are involved in multiple physiological and pathological phenomenon by acting as cross-linkers between actin cytoskeleton and plasma membrane in polarized cells [4][5][6][7] . As the founding member of the ERM, ezrin is essential for cell migration 8 , inflammatory response regulation 9 , villus morphogenesis 10 and gastric acid secretion [11][12][13][14] . It has also been proved to be an important marker for cancer metastasis 15,16 .…”

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confidence: 99%

Celastrol inhibits ezrin-mediated migration of hepatocellular carcinoma cells

Song

et al. 2020

Sci Rep

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Chemicals and antibody. Celastrol was ordered from MedChemExpress under the item number HY-13067. Antibodies against ezrin, ezrin pT567 and α-tubulin (DM1A) were purchased from Cell Signaling Technology. Antibodies against ROCK2 and ROCK2 (phospho S1366) were purchased from Abcam. Anti-GFP antibodies were purchased from Santa Cruz Biotechnology. Anti-FLAG-tag (M2) antibody was from Sigma. plasmids. The development of bacterial expression vectors containing human ezrin fused to histidine was described previously 61. EGFP-tagged ezrin plasmids was produced as described before 62. PCR amplified ROCK2 cDNA was cloned into the 3× FLAG-Myc-CMV-24 vector (Sigma) by BamHI and RcoRI digestion. The mutanted of ezrin and ROCK2 were constructed by Fast Mutagenesis Kit (Vazyme Biotech). All plasmids were verified by sequencing (Tsingke Biological Tech). cell culture and transfection. MHCC97H was a gift from Professor Yong Chen (Fourth Military Medical University) 20. HepG2 and HEK293T cells were from American Type Culture Collection (ATCC). Huh7 cells were from National Infrastructure of Cell Line Resource. The cells were incubated according to ATCC instruction. Cells were transfected using conventional calcium phosphate method or the Lipofectamine 3,000 (Invitrogen) according to manufacturer's instructions. Determination of cell viability (MTS assay). Celastrol cytotoxicity was assessed with the use of a CellTiter 96 AQueous One Solution Cell Proliferation Assay (Promega), which is a form of the 3-(4,5-dimethylthiazol-2-yl)-5-(3-carboxymethoxyphenyl)-2-(4-sulfophenyl)-2H-tetrazolium inner salt (MTS) assay. Cells (5,000 cells/well) were plated in 96-well plates, treated with various concentrations of celastrol for 24 h. After celastrol treatment, cell viability was determined by adding a small amount of the One Solution Reagent directly to culture wells, incubating for 3 h and then recording the absorbance at 490 nm with a 96-well plate reader. Wound healing assay. MHCC97H, HepG2 or Huh7 cells with logarithmic growth phases, was cultivated in 12 orifices with 5 × 10 5 cells density. Each group has three replicates. After a starvation of 6 h in serum-free medium, 10 μl Tip was taken to vertical scratch "#" glyph at the bottom of the culture plate. The floating cells were washed off with PBS and the celastrol was dissolved in medium supplemented with 20% FBS. Samples were photographed at 0 h, 4 h and 8 h. Image J software was used to measure and compare the effects of celastrol on cell migration. Single cell tracking assay. The cell migration by single cell tracking assay was performed as previously described 32. Briefly, MHCC97H cells were laid in a Silian dish. After stabilized, the cells were starved with Opti-MEM for 6 h, and then cultured in DMEM containing 20% FBS. The movement track of 10 single-cells in each field of view were recorded by microscope for 6 h, with 10 min interval at once.

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“…Light-sheet fluorescence microscopy allows rapid acquisition of three-dimensional images over large fields of view and over long durations (103). Three-dimensional images are compiled from successive light-sheet illuminations of thin, two-dimensional optical sections.…”

Section: Modeling Membrane Organelle Dynamics and Cell Fate In Organoidsmentioning

confidence: 99%

Phase separation drives decision making in cell division

Liu

Wang

et al. 2020

Journal of Biological Chemistry

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Liquid-liquid phase separation (LLPS) of biomolecules drives the formation of subcellular compartments with distinct physicochemical properties. These compartments, free of lipid bilayers and therefore called membraneless organelles, include nucleoli, centrosomes, heterochromatin, and centromeres. These have emerged as a new paradigm to account for subcellular organization and cell fate decisions. Here we summarize recent studies linking LLPS to mitotic spindle, heterochromatin, and centromere assembly and their plasticity controls in the context of the cell division cycle, highlighting a functional role for phase behavior and material properties of proteins assembled onto heterochromatin, centromeres, and central spindles via LLPS. The techniques and tools for visualizing and harnessing membraneless organelle dynamics and plasticity in mitosis are also discussed, as is the potential for these discoveries to promote new research directions for investigating chromosome dynamics, plasticity, and inter-chromosome interactions in the decision-making process during mitosis.

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Gastric Parietal Cell Physiology and Helicobacter pylori–Induced Disease

Cited by 81 publications

References 162 publications

NDP52 tunes cortical actin interaction with astral microtubules for accurate spindle orientation

NDP52 tunes cortical actin interaction with astral microtubules for accurate spindle orientation

Celastrol inhibits ezrin-mediated migration of hepatocellular carcinoma cells

Phase separation drives decision making in cell division

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