• Title/Summary/Keyword: mRNA expression

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Effects of the Repression of sphpr1 Expression on Growth and mRNA Export in Fission Yeast (분열효모에서 sphpr1 유전자의 결실이 생장 및 mRNA Export에 미치는 영향)

  • Lee, Hyun-Joo;Yoon, Jin-Ho
    • Korean Journal of Microbiology
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    • v.48 no.2
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    • pp.171-174
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    • 2012
  • THOC1/Hpr1 is one subunit of THO complex that is an evolutionally conserved assembly involved in the mRNP packaging and mRNA export during transcription elongation. In fission yeast Schizosaccharomyces pombe, an ortholog (spHpr1) of THOC1/Hpr1 was identified based on sequence alignment. A deletion mutant in a diploid strain was constructed by replacing one of spHpr1-coding region with a $kan^r$ gene using one-step gene disruption method. Tetrad analysis showed that the sphpr1 is essential for growth. Over-expression of sphpr1 from strong nmt1 promoter caused no defects of growth and mRNA export. However, repression of the sphpr1 expression resulted in growth inhibition accompanied by accumulation of poly$(A)^+$ RNA in the nucleus. These results suggest that spHpr1 is involved in mRNA export from the nucleus to cytoplasm.

Polymorphism of the Promoter Region of Hsp70 Gene and Its Relationship with the Expression of HSP70mRNA, HSF1mRNA, Bcl-2mrna and Bax-AMrna in Lymphocytes in Peripheral Blood of Heat Shocked Dairy Cows

  • Cai, Yafei;Liu, Qinghua;Xing, Guangdong;Zhou, Lei;Yang, Yuanyuan;Zhang, Lijun;Li, Jing;Wang, Genlin
    • Asian-Australasian Journal of Animal Sciences
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    • v.18 no.5
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    • pp.734-740
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    • 2005
  • The blood samples were collected from dairy cows at the same milking stage. The single-strand conformation polymorphism (PCR-SSCP) method was used to analyze for polymorphism at the 5'flanking region of the hsp70 gene. The mRNA expression levels of HSP70, HSF1, Bcl-2 and Bax-$\alpha$ at different daily-mean-temperature were analyzed by relative quantitative RTPCR. The DNA content, cell phase and the ratio of apoptosis of lymphocytes in peripheral blood of dairy cattle at different daily-meantemperature were determined by FCM. The PCR-SSCP products of primer pair 1 showed polymorphisms and could be divided into four genotypes: aa, ab, ac, cc, with the cis-acting element (CCAAT box) included. Mutations in the hsp70 5'flanking region (468-752 bp) had different effects on mRNA expression of HSP70, HSF1, Bcl-2 and Bax-$\alpha$. The ac genotypic cows showed higher expressions of HSP70mRNA, HSF1mRNA and Bcl-2mRNA/Bax-$\alpha$mRNA and lower ratio of apoptosis. These mutation sites can be used as molecular genetic markers to assist selection for anti-heat stress cows.

Effects of 3-methylcholanthrene on the Expression of Drug Metabolizing Enzyme Genes in Monkey Brain (원숭이 뇌 약물대사효소 유전자 발현에 미치는 3-methylcholanthrene 영향)

  • 이경원;아사오카;신윤용
    • Environmental Mutagens and Carcinogens
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    • v.24 no.1
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    • pp.40-45
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    • 2004
  • In order to understand the mechanism of the regulation of drug metabolizing enzyme gene expression, we have studied the induction of CYP1A1 and GSTα, μ, π enzymes in Japanese monkey and rhesus monkey after the treatment with 3-methylcholanthrene (3MC) and di-n- butyl phthalate (DBP) and bisphenol A (BPA). The levels of mRNA were measured_by RT-PCR in brain, intestine and liver. In the case of adult monkey, treatment with 3MC induced CYP1A1 mRNA in brain by 2-fold. The treatment with DBP induced CYP1A1 mRNA. Effects of 3MC and DBP on GST mRNA expression was not clear. But GSTμ was slightly inhibited by the treatment with 3MC and DBP. GSTα was not induced by the treatment with 3MC and DBP in brain. GSTπ was slightly induced by the treatment with 3MC and DBP in brain. In the case of fetus monkey, the basal levels of fetus CYP1A1 mRNA and GSTs mRNA were relatively low compared to adult monkey. As the age of monkey increased, the basal levels of CYP1A1 mRNA were also increased. 3MC induced the expression of CYP1A1 mRNA in liver, whereas it didn't significantly induce CYP1A1 mRNA in brain. The levels of GSTμ and GSTα were not changed by the treatment with 3MC and DBP. GSTπ was slightly induced by the treatment with 3MC and DBP.

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Effects of 3-methylcholanthrene on the Expression of Drug Metabolizing Enzyme Genes in Monkey Intestine (원숭이 소장 약물대사효소 유전자 발현에 미치는 3-methylcholanthrene 영향)

  • 이경원;아사오카;신윤용
    • Environmental Mutagens and Carcinogens
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    • v.24 no.1
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    • pp.19-24
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    • 2004
  • In order to understand the mechanism of the regulation of drug metabolizing enzyme gene expression, we have studied the induction of CYP1A1 and $GST\alpha,$ $\mu,$ $\pi$ enzymes in Japanese monkey and rhesus monkey after the treatment with 3-methylcholanthrene (3MC) and di-n-butyl phthalate (DBP) and bisphenol A (BPA). The levels of mRNA were measured by RT-PCR in brain, intestine and liver. In the case of adult monkey, treatment with 3MC induced CYP1A1 mRNA in intestine by 11-fold. The treatment with DBP induced CYP1A1 mRNA. Effects of 3MC and DBP on GST mRNA expression was not clear. But $GST\mu$ was slightly inhibited by the treatment with 3MC and DBP. $GST\alpha$ was induced in intestine by 1.5-fold. $GST\pi$ was slightly induced by the treatment with 3MC and DBP in intestine. In the case of fetus monkey, the basal levels of fetus CYP1A1 mRNA and GSTs mRNA were relatively low compared to adult monkey. As the age of monkey increased, the basal levels of CYP1A1 mRNA were also increased. 3MC induced the expression of CYP1A1 mRNA didn't significantly induce CYP1A1 mRNA in intestine. The levels of $GST\mu$ and $GST\pi$ were not changed by the treatment with 3MC and DBP. $GST\pi$ was slightly induced by the treatment with 3MC and DBP.

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siRNA-mediated Inhibition of hTERC Enhances Radiosensitivity of Cervical Cancer

  • Chen, Min;Xing, Li-Na
    • Asian Pacific Journal of Cancer Prevention
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    • v.13 no.12
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    • pp.5975-5979
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    • 2012
  • Background: To investigate the influence of telomerase activity, apoptosis, radiosensitivity of cervical cancer after siRNA-mediated knockdown of telomerase RNA and evaluate in vivo growth with gene interference. Methods: We studied siRNA-targeting-telomerase RNA transfection into the Hela cell line. Expression of hTERC mRNA was detected by RT-PCR and telomerase activity was measured by the TRAP assay. Growth inhibition was determined by MTT assay and radiosensitivity of the cervical cancer cells was examined by colony formation assay. In addtion, effects of hTERC inhibition in vivo were studied by injection of siRNA-transfected Hela cells into nude mice. Results: The hTERC siRNA effectively downregulated the expression of hTERC mRNA and also reduced the telomerase activity to 30% of the untreated control vlaue. The viability of hTERC siRNA transfected Hela cells was reduced by 44.7% after transfection. After radiation treatment, the radiosensitivity of Hela cells with hTERC knockdown was increased. In vivo, the tumors developing from the hTERC siRNA-transfected cells were of reduced size, indicating that the hTERT siRNA also depressed the tumorigenic potential of the Hela cells. Conclusions: Our results supported the concept of siRNA-mediated inhibition of telomerase mRNA which could inhibit the expression of hTERC and telomerase activity. Furthermore, radiosensitivity was upregulated after knockdown the hTERC in vivo and in vitro.

Effects of spNab2 Deletion and Over-Expression on mRNA Export (분열효모에서 spNab2 유전자의 결실돌연변이 및 과발현에 대한 분석)

  • Yoon, Jin-Ho
    • Korean Journal of Microbiology
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    • v.45 no.4
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    • pp.300-305
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    • 2009
  • We constructed the deletion mutants of fission yeast Schizosaccharomyces pombe spNab2 gene that is homologous to poly(A)-binding protein NAB2 in budding yeast Saccharomyces cerevisiae, which plays crucial roles in mRNA 3' end formation and mRNA export from nucleus into the cytoplasm. A null mutant in an $h^+$/ $h^+$ diploid strain was constructed by replacing the spNab2-coding region with an $ura4^+$ gene using one-step gene disruption method. Tetrad analysis showed that the spNab2 is not essential for vegetative growth and mRNA export. However, over-expression of spNab2 cause the severe growth defects and intensive accumulation of poly(A) RNA in the nucleus. Also, the spNab2-GFP fusions were localized mainly in the nucleus. These results suggest that spNab2 is also involved in mRNA export out of the nucleus.

Tissue- and Reproductive Organ-specific Expression of Protease Nexin-1 in Sprague-Dawley Rat (흰쥐에서 단백질 분해효소 저해제, Nexin-1의 조직 및 생식기관 특이적 유전자 발현)

  • 고정재;김남근;김진규;최명진;정형민;서승염;김윤희;이현환;차광열
    • Development and Reproduction
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    • v.2 no.2
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    • pp.135-140
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    • 1998
  • Protease Nexin-1 (PN-1) inhibits the activity of several serine proteases including thrombin, urokinase (uPA)-type plasminogen activator and trypsin. Tissue- and reproductive organ-specific mRNA levels of the PN-1 were investigated in Sprague-Dawley adult rat. PN-1 mRNA expression in rats was found in brain (forebrain, hindbrain), heart, liver, lung, ovary and oviduct. The level of PN-1 mRNA in male and female among the tissues was the highest in forebrain of the female. PN-1 expression in reproductive organs was found only in ovary and oviduct. These results suggest that PN-1 expression is dependent on the sex and may be related to folliculogenesis and early embryogenesis.

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Ritanserin, a 5HT2/1C Receptor Antagonist, Does Not Block Cocaine-Induced Behavioral Alterations and zif268 mRNA Expression in the Striatum of the Rats

  • Choe, Eun-Sang;Kim, Jong-Yeon
    • The Korean Journal of Physiology and Pharmacology
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    • v.4 no.5
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    • pp.355-359
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    • 2000
  • Cocaine induces immediate early gene expression and behavioral changes by blocking dopamine transporters in the terminals of nigrostriatal neurons in the striatum. The pharmacological role of serotonin 2/1C (5HT2/1C) receptors in cocaine-induced expression of zif268 (NGFI-A, egr1 and Krox-24) mRNA, a member of the zinc finger, was investigated using quantitative in situ hybridization histochemistry in vivo. Behavioral alterations induced by cocaine were also monitored in relation with blockade of the receptors. Systemic injection of ritanserin (1 mg/kg, s.c.), a 5HT2/1C receptor antagonist, did not reverse behavioral alterations and zif268 mRNA gene expression induced by 15 mg/kg cocaine, i.p., in the dorsal and ventral striatum. These data indicate that ritanserin-sensitive 5HT2/1C receptors are not necessary for cocaine-induced behavioral alterations and zif268 mRNA gene expression in the striatum.

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Skin Wound Healing Effects and Action Mechanism of Acai Berry Water Extracts

  • Kang, Mi Hyun;Choi, Seunghye;Kim, Bae-Hwan
    • Toxicological Research
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    • v.33 no.2
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    • pp.149-156
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    • 2017
  • The purpose of this study was to investigate the wound healing effect of acai berry water extracts (ABWE) and a possible underlying mechanism involved in its action using various in vitro and in vivo models. The wound healing effect of ABWE was evaluated by migration assay using HS68 fibroblast cells. In addition, its effect on mRNA expression of procollagen, fibronectin, and MMP-1 was determined. Moreover, the wound healing effect of ABWE was evaluated in in vivo wound models through macroscopic and microscopic observation. In addition, mRNA expression levels of wound related genes were determined. Results revealed that ABWE was not cytotoxic. It increased migration of HS68 fibroblast cells. ABWE increased mRNA expression levels of fibronectin but decreased the mRNA expression levels of MMP-1. ABWE also showed significantly potent wound healing effect in vivo based on macroscopic and histopathological observation and mRNA expression evaluation for wound related genes. Taken together, our results indicated that ABWE might have potential as a wound healing agent.

Ceramide-Mediated c-jun Gene Expression in U-937 Cells (U-937 세포에 있어서 세라마이드에 의한 c-jun 유전자 발현의 조절)

  • Kim, Won-Ho;Kim, Mie-Young;Choi, Kyung-Hee
    • YAKHAK HOEJI
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    • v.41 no.1
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    • pp.81-85
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    • 1997
  • Ceramide has been suggested as an important mediator of the effects of extracellular agonists on cell growth inhibition, differentiation, apoptosis. However the biochemical sign aling mechanism involved in transducing the effects of ceramide on leukemia cell differentiation is still unclear. In these respects, we examined the regulatory effects of ceramide on c-jun gene expression during differentiation. In U-937 cells. ceramide increased c-jun mRNA levels in a time-dependent manner. The half life, of c-jun mRNA was 30 min. In contrast, inhibition of protein synthesis with cycloheximide in the absence, of transcription with actinomycin D increased the half-life of c-jun mRNA in ceramide-treated U-937 cells to more than 90 min. In order to examine whether ceramide-inhibited c-jun gene expression is regulated through ceramide-activated protein phosphatase (CAPP), a direct target for the action of ceramide, okadaic acid were treated to the cells. Okadaic acid inhibited enhancement of c-jun mRNA induced by C2-ceramide in a dose-dependent manner. These results suggested that ceramide increases c-jun mRNA level during differentiation in U-937 cells and regulates the gene expression on posttranscriptional level. In addition, we provide the evidence that CAPP is involved in ceramide-induced c-jun gene expression in U-937 cells.

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