• Title/Summary/Keyword: RSK

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Effect of Feeding Rubber Seed Kernel and Palm Kernel Cake in Combination on Nutrient Utilization, Rumen Fermentation Characteristics, and Microbial Populations in Goats Fed on Briachiaria humidicola Hay-based Diets

  • Chanjula, P.;Siriwathananukul, Y.;Lawpetchara, A.
    • Asian-Australasian Journal of Animal Sciences
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    • v.24 no.1
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    • pp.73-81
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    • 2011
  • Six male crossbred (Thai Native${\times}$Anglo Nubian) goats, with an average initial weight of $22{\pm}2\;kg$, were randomly assigned according to a $3{\times}2$ factorial arrangement in a $6{\times}6$ Latin square design with a 21-d period to evaluate the effect of feeding rubber seed kernel (RSK) and palm kernel cake (PKC) in combination on nutrient utilization, rumen fermentation characteristics, and nitrogen utilization. The dietary treatments were as follows: i) concentrate containing 0% RSK and 20% PKC ($T_1$), ii) 0% RSK and 30% PKC ($T_2$), iii) 20% RSK and 20% PKC ($T_3$), iv) 20% RSK and 30% PKC ($T_4$), v) 30% RSK and 20% PKC ($T_5$), and vi) 30% RSK and 30% PKC ($T_6$). During the experiment, signal hay was given on an ad libitum basis as the roughage. It was found that RSK levels and PKC levels had no interaction effects on feed intake, apparent digestibility, $NH_3$-N, blood metabolites, VFA concentrations, and nitrogen utilization, but there were interactions between RSK levels and PKC levels with respect to total DMI (kg/d) and total VFA concentrations, and goats receiving 30% RSK had lower values (p<0.05) than those receiving 0 and 20% RSK, respectively. Feeding different PKC levels did not affect (p>0.05) feed intake, digestibility, rumen fermentation patterns, blood metabolites, and nitrogen utilization. However, increasing RSK levels (>20%) resulted in a slightly lower daily DMI (% BW and g/kg $BW^{0.75}$), apparent digestibility (NDF and ADF), total N intake, and N excretion than in goats fed on 0 and 20% RSK. BUN, blood glucose, and propionate were variable among treatment and were highest in 0% RSK with the 20% PKC fed group having values which were higher than those in other groups. However, there were no differences (p>0.05) among treatments with respect to N retention, PD output, and microbial N supply. Based on this study, RSK levels up to 20% and PKC at 20-30% in concentrate could be efficiently utilized for goats fed on signal hay.

The p90rsk-mediated signaling of ethanol-induced cell proliferation in HepG2 cell line

  • Kim, Han Sang;Kim, Su-Jin;Bae, Jinhyung;Wang, Yiyi;Park, Sun Young;Min, Young Sil;Je, Hyun Dong;Sohn, Uy Dong
    • The Korean Journal of Physiology and Pharmacology
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    • v.20 no.6
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    • pp.595-603
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    • 2016
  • Ribosomal S6 kinase is a family of serine/threonine protein kinases involved in the regulation of cell viability. There are two subfamilies of ribosomal s6 kinase, (p90rsk, p70rsk). Especially, p90rsk is known to be an important downstream kinase of p44/42 MAPK. We investigated the role of p90rsk on ethanol-induced cell proliferation of HepG2 cells. HepG2 cells were treated with 10~50 mM of ethanol with or without ERK and p90rsk inhibitors. Cell viability was measured by MTT assay. The expression of pERK1, NHE1 was measured by Western blots. The phosphorylation of p90rsk was measured by ELISA kits. The expression of Bcl-2 was measured by qRT-PCR. When the cells were treated with 10~30 mM of ethanol for 24 hour, it showed significant increase in cell viability versus control group. Besides, 10~30 mM of ethanol induced increased expression of pERK1, p-p90rsk, NHE1 and Bcl-2. Moreover treatment of p90rsk inhibitor attenuated the ethanol-induced increase in cell viability and NHE1 and Bcl-2 expression. In summary, these results suggest that p90rsk, a downstream kinase of ERK, plays a stimulatory role on ethanol-induced hepatocellular carcinoma progression by activating anti-apoptotic factor Bcl-2 and NHE1 known to regulate cell survival.

Inhibition of p90RSK activation sensitizes triple-negative breast cancer cells to cisplatin by inhibiting proliferation, migration and EMT

  • Jin, Yujin;Huynh, Diem Thi Ngoc;Kang, Keon Wook;Myung, Chang-Seon;Heo, Kyung-Sun
    • BMB Reports
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    • v.52 no.12
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    • pp.706-711
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    • 2019
  • Cisplatin (Cis-DDP) is one of the most widely used anti-cancer drugs. It is applicable to many types of cancer, including lung, bladder, and breast cancer. However, its use is now limited because of drug resistance. p90 ribosomal S6 kinase (p90RSK) is one of the downstream effectors in the extracellular signal-regulated protein kinases 1 and 2 (ERK1/2) pathway and high expression of p90RSK is observed in human breast cancer tissues. Therefore, we investigated the role of p90RSK in the Cis-DDP resistance-related signaling pathway and epithelial-mesenchymal transition (EMT) in breast cancer cells. First, we discovered that MDA-MB-231 cells exhibited more Cis-DDP resistance than other breast cancer cells, including MCF-7 and BT549 cells. Cis-DDP increased p90RSK activation, whereas the inactivation of p90RSK using a small interfering RNA (siRNA) or dominant-negative kinase mutant plasmid overexpression significantly reduced Cis-DDP-induced cell proliferation and migration via the inhibition of matrix metallopeptidase (MMP)2 and MMP9 in MDA-MB-231 cells. In addition, p90RSK activation was involved in EMT via the upregulation of mRNA expression, including that of Snail, Twist, ZEB1, N-cadherin, and vimentin. We also investigated NF-κB, the upstream regulator of EMT markers, and discovered that Cis-DDP treatment led to NF-κB translocation in the nucleus as well as its promoter activity. Our results suggest that targeting p90RSK would be a good strategy to increase Cis-DDP sensitivity in triple-negative breast cancers.

Actin Dysfunction Induces Cell Cycle Delay at G2/M with Sustained ERK and RSK Activation in IMR-90 Normal Human Fibroblasts

  • Shrestha, Deepmala;Choi, Daeun;Song, Kiwon
    • Molecules and Cells
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    • v.41 no.5
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    • pp.436-443
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    • 2018
  • The actin cytoskeleton plays a key role in the entry of mitosis as well as in cytokinesis. In a previous study, we showed that actin disruption delays mitotic entry at G2/M by sustained activation of extracellular signal-related kinase 1/2 (ERK1/2) in primary cells but not in transformed cancer cell lines. Here, we examined the mechanism of cell cycle delay at G2/M by actin dysfunction in IMR-90 normal human fibroblasts. We observed that de-polymerization of actin with cytochalasin D (CD) constitutively activated ribosomal S6 kinase (RSK) and induced inhibitory phosphorylation of Cdc2 (Tyr 15) in IMR-90 cells. In the presence of an actin defect in IMR-90 cells, activating phosphorylation of Wee1 kinase (Ser 642) and inhibitory phosphorylation of Cdc25C (Ser 216) was also maintained. However, when kinase-dead RSK (DN-RSK) was overexpressed, we observed sustained activation of ERK1/2, but no delay in the G2/M transition, demonstrating that RSK functions downstream of ERK in cell cycle delay by actin dysfunction. In DN-RSK overexpressing IMR-90 cells treated with CD, phosphorylation of Cdc25C (Ser 216) was blocked and phosphorylation of Cdc2 (Tyr 15) was decreased, but the phosphorylation of Wee1 (Ser 642) was maintained, demonstrating that RSK directly controls phosphorylation of Cdc25C (Ser 216), but not the activity of Wee1. These results strongly suggest that actin dysfunction in primary cells activates ERK1/2 to inhibit Cdc2, delaying the cell cycle at G2/M by activating downstream RSK, which phosphorylates and blocks Cdc25C, and by directly activating Wee1.

Phosphorylation of REPS1 at Ser709 by RSK attenuates the recycling of transferrin receptor

  • Kim, Seong Heon;Cho, Jin-hwa;Park, Bi-Oh;Park, Byoung Chul;Kim, Jeong-Hoon;Park, Sung Goo;Kim, Sunhong
    • BMB Reports
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    • v.54 no.5
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    • pp.272-277
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    • 2021
  • RalBP1 associated EPS domain containing 1 (REPS1) is conserved from Drosophila to humans and implicated in the endocytic system. However, an exact role of REPS1 remains largely unknown. Here, we demonstrated that mitogen activated protein kinase kinase (MEK)-p90 ribosomal S6 Kinase (RSK) signaling pathway directly phosphorylated REPS1 at Ser709 upon stimulation by epidermal growth factor (EGF) and amino acid. While REPS2 is known to be involved in the endocytosis of EGF receptor (EGFR), REPS1 knockout (KO) cells did not show any defect in the endocytosis of EGFR. However, in the REPS1 KO cells and the KO cells reconstituted with a non-phosphorylatable REPS1 (REPS1 S709A), the recycling of transferrin receptor (TfR) was attenuated compared to the cells reconstituted with wild type REPS1. Collectively, we suggested that the phosphorylation of REPS1 at S709 by RSK may have a role of the trafficking of TfR.

Sustained Intracellular Acidosis Triggers the Na+/H+ Exchager-1 Activation in Glutamate Excitotoxicity

  • Lee, Bo Kyung;Jung, Yi-Sook
    • Biomolecules & Therapeutics
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    • v.25 no.6
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    • pp.593-598
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    • 2017
  • The $Na^+/H^+$ exchanger-1 (NHE-1) is a ubiquitously expressed pH-regulatory membrane protein that functions in the brain, heart, and other organs. It is increased by intracellular acidosis through the interaction of intracellular $H^+$ with an allosteric modifier site in the transport domain. In the previous study, we reported that glutamate-induced NHE-1 phosphorylation mediated by activation of protein kinase C-${\beta}$ (PKC-${\beta}$) in cultured neuron cells via extracellular signal-regulated kinases (ERK)/p90 ribosomal s6 kinases (p90RSK) pathway results in NHE-1 activation. However, whether glutamate stimulates NHE-1 activity solely by the allosteric mechanism remains elusive. Cultured primary cortical neuronal cells were subjected to intracellular acidosis by exposure to $100{\mu}M$ glutamate or 20 mM $NH_4Cl$. After the desired duration of intracellular acidosis, the phosphorylation and activation of PKC-${\beta}$, ERK1/2 and p90RSK were determined by Western blotting. We investigated whether the duration of intracellular acidosis is controlled by glutamate exposure time. The NHE-1 activation increased while intracellular acidosis sustained for >3 min. To determine if sustained intracellular acidosis induced NHE-1 phosphorylation, we examined phosphorylation of NHE-1 induced by intracellular acidosis by transient exposure to $NH_4Cl$. Sustained intracellular acidosis led to activation and phosphorylation of NHE-1. In addition, sustained intracellular acidosis also activated the PKC-${\beta}$, ERK1/2, and p90RSK in neuronal cells. We conclude that glutamate stimulates NHE-1 activity through sustained intracellular acidosis, which mediates NHE-1 phosphorylation regulated by PKC-${\beta}$/ERK1/2/p90RSK pathway in neuronal cells.

Taxonomic Redescription of Loxophyllum perihoplophorum and L. rostratum (Ciliophora: Pleurostomatida) from Korea

  • Kim, Se-Joo;Min, Gi-Sik
    • Animal Systematics, Evolution and Diversity
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    • v.31 no.4
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    • pp.277-283
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    • 2015
  • Two pleurostomatid ciliates, Loxophyllum perihoplophorum Buddenbrock, 1920 and L. rostratum Cohn, 1866, were collected from the coastal waters of the East Sea, Korea. Their morphologies are described based on live observation and protargol staining, and morphometrics are provided. Loxophyllum perihoplophorum is characterized by the following features: 200-650 μm long in vivo; body slender leaf-shaped, flexible and contractile, with thin and wide extrusome-belted zone; 2 macronuclear nodules (Ma) and 1 micronucleus (Mi); 7-9 contractile vacuoles (CV) positioned along dorsal margin; extrusomes (Ex) evenly distributed along edge of entire body, with about 10 dorsal warts (Wa); 9-11 left (LSK) and 19-22 right somatic kineties (RSK), 4-5 furrows (Fu) on left side. Loxophyllum rostratum is about 100-130 μm long in vivo; body oblate leaf-shaped, contractile, convex ventral side and S-shaped dorsal side, beak-like anterior end; 2 Ma and 1 Mi; 1 CV terminally located; Ex distributed along edge of entire body, with about 9-10 dorsal Wa; 7-8 LSK and 15-19 RSK, ca. 5 Fu on left body side. In addition, sequences of small subunit ribosomal DNA were determined from these two Loxophyllum species and compared with the known Loxophyllum sequences.

Taxonomic Study of Poorly-known Marine Pleurostomatid Ciliates of Litonotus paracygnus and L. pictus (Ciliophora: Pleurostomatida) from Korea

  • Kim, Se-Joo;Min, Gi-Sik
    • Animal Systematics, Evolution and Diversity
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    • v.25 no.2
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    • pp.167-178
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    • 2009
  • Two poorly known and often confused pleurostomatid ciliates, Litonotus paracygnus Song, 1994 and L. pictus Gruber, 1884, were collected from the coastal waters of Yeonggeumjeong and Bongpo-port, Gangwondo in the East Sea and from the Iwon tide embankment near Ganwol-do, Chungcheongnam-do in the Yellow Sea, Korea. These species were described based on live observations, the protargol-impregnation and morphometrics of the species. Also provided are their diagnoses. The small subunit ribosomal DNA (SSU rDNA) sequences of these species were compared with previously known sequences of related species. The diagnostics of the two Litonotus species are as follows. L. paracygnus: 150-300 $\mu$m long in vivo, strongly contractile neck region, two ellipsoid macronuclei (Ma) and one micronucleus (Mi), 7 left (LSK) and 11-14 right somatic kineties (RSK), 2-4 contractile vacuoles (CV) located on the posterior end, extrusemes (Ex) distributed on the anterior region of the ventral margin only. L. pictus: about 200-600 $\mu$m long in vivo, extremely contractile, beautiful body color with rows of yellow to yellow-brownish cortical pigment granules, 12-21 Ma arranged in moniliform pattern, infrequently vermiform, 7-11 LSK and 18-26 RSK, several CV located on both margins, Ex distributed on the anterior region of the ventral margin only. In this study, this genus was firstly recorded in Korea.

3,4-Dihydroxytoluene suppresses UVB-induced wrinkle formation by inhibiting Raf-1

  • Park, Sang-Hee;Kang, Nam Joo
    • Korean Journal of Food Science and Technology
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    • v.52 no.4
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    • pp.385-395
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    • 2020
  • This study examined the effect of 3,4-dihydroxytoluene (DHT) on UVB-induced photoaging and determined its molecular mechanisms, using HaCaT human keratinocytes and SKH-1 hairless mice. DHT suppressed UVB-induced matrix metalloproteinase-1 (MMP-1) expression in HaCaT cells. In vivo data from mouse skin supported that DHT decreased UVB-induced wrinkle formation, epidermal thickness, and matrix metalloproteinase-13 (MMP-13) expression. DHT appeared to exert its anti-aging effects by suppressing UVB-induced Raf-1 kinase activity and subsequent attenuation of UVB-induced phosphorylation of MEK, ERK, and p90RSK in HaCaT cells. In vitro and in vivo pull-down assays revealed that DHT bound with Raf-1 in ATP-noncompetitive manner. Overall, DHT appears to anti-photoaging effects in vitro and in vivo through the suppression of Raf-1 kinase activity and may have potential as a treatment for the prevention of skin aging.