12-O-Tetradecanoylphorbol-13-Acetate에 의한 HL-60 세포 분화유도중 Vimentin 유전자 전사조절에 대한 AP-1의 역할

임규,김진희,권도원,김승민,이명선,윤경아,손미영,박종일,윤완희,황병두 충남대학교 생물공학연구소 1999 생물공학연구지 Vol.7 No.-

Purpose: To gain insight on the role of AP-1 in transcriptional regulation of vimentin gene during differentiation of HL-60 cells by 12-O-tetradecanoylphorbol-13-acetate (TPA), the levels of vimentin mRNA and AP-1 have been investigated with Northern blot hybridization and DNA mobility shift assay. Materials and Methods: HL-60 cells were grown in RPMI 1640 medium supplemented with 10% heat-inactivated fetal calf serum and antibiotics in a humidified 5% CO_2 at 37℃. Total RNA was prepared by a modification of the method of Karlinsey et al. Northern blot hybridization was performed by the method of Virca et al. EcoRI fragment of pVIM-GEM was used as probe for vimentin mRNA. DNA mobility shift assay was performed by the method of Lim et al. End labeled DNA probe(Upper strand, 5'-CGCTTGATGAGTCAGCCG- 3') for AP-1 binding activity was mixed with nuclear extracts in a 20 μl reaction volume containing 300 mM KCI, 60 mM HEPES, pH 7.9, 25 mM MgCI_2, 1 mM EDTA, 1 mM DTT, 60% glycerol, and 2 ㎍ of poly[dI-dC]. Results: TPA increased vimentin mRNA levels, with maximal stimulation reached at 24 hr. The level of vimentin mRNA was induced in proportion to the concentration of TPA. TPA-induced vimentin mRNA was almost reduced by actinomycin-D pretreatment. TPA-induced stimulation of vimentin gene was completely reduced by staurosporin pretreatment. In DNA mobility shift assay, AP-1 newly appeared at 24 hr during TPA-induced differentiation and was almost not detected after the pretreatment of staurosporin. Conclusions: These results suggest that the induction of vimentin mRNA during TPA-dependent differentiation in HL-60 cells may be mediated by protein kinases C signal transduction and AP-1 is important to transcriptional regulation.

백서 간재생중 H2B Histone 유전자 발현에 대한 Octamer Binding Transcription Factor의 역할

임규,방재웅,최두식,손미영,이영철,박청,김태동,김영래,박종일,윤완희,박승길,황병두 한국노화학회 2001 한국노화학회지 Vol.11 No.1

일차배양 Sertoli Cell에서 Testosterone에 의한 세포성 핵암유전자 발현에 관한 연구

임규,유지헌,김계영,권기량,황병두,Lim, Kyu,Yoo, Ji-Hun,Kim, Kye-Young,Kweon, Gi-Ryang,Hwang, Byung-Doo 생화학분자생물학회 1993 한국생화학회지 Vol.26 No.1

일차배양 Sertoli cell에서 testosterone에 의한 c-myc 등 세포성 핵암유전자 발현 및 그 조절기전을 검색하여 다음과 같은 결과를 얻었다. Steady state에서 발현되는 Sertoli cell 세포성 핵암유전자는 c-fos, c-myc, c-jun이었으며 c-myb은 검출되지 않았고 이때 mRNA 크기는 각각 2.2 kb, 2.4 kb, 2.7 kb에 해당되었다. Testosterone에 의한 c-myc 및 c-jun mRNA의 발현은 시간경과에 따라 증가하였으며 유도되는 시간은 모두 16시간에 최고치에 달하여 유사하였다. c-Myc은 testosterone 농도에 비례해서 그리고 cycloheximide 처리로 각각 전사가 증가되었으나 actinomycin D 처리로 억제되었다. 연령에 따른 Sertoli cell에서의 c-myc mRNA 발현에 대한 testosterone의 영향은 steady state에서는 8일령이 가장 높았으며, testosterone에 의한 유도는 8일령, 14일령에서는 나타났으나 28일령에서는 유도되지 않았다. 이상의 결과로 미루어 보아 testosterone에 의해 유도된 c-myc mRNA가 testosterone에 의해 조절되는 Sertoli cell의 다른 유전자 발현의 조절에 관련되어 있을 것이라 시사된다. The expression of the nuclear protooncogenes have been associated with the cell proliferation and differentiation into changes in nuclear function. To evaluate the possibility that the nuclear protooncogenes play roles testosterone-dependent gene regulation, the effects of testosterone on the expression of the nuclear protooncogenes have been investigated in primary Sertoli cell cultures. c-Myc, c-jun and c-fos were expressed in steady state of primary Sertoli cells but c-myb was not detected. Testosterone increased c-myc and c-jun mRNA with maximal stimulation reached in 16 h. The induction of c-myc was dependent on the concentrations of testosterone. The testsoterone induced c-myc mRNA level was also increased in the cells treated with cycloheximide, but reduced by actinomycin D pretreatment. Even in the absence of testosterone, c-myc mRNA was clearly detectable in Sertoli cells from 8-days-old rats, but hardly detectable in cells from 14 and 28 days of age. Testosterone stimulated c-myc expression in the Sertoli cells from 8 and 14-days-old rats. These results suggest that transient expression of c-myc may play some roles to an intermediate step required for testosterone-dependent regulation of other genes expression of Sertoli cells.

HL-60 세포에서 Myeloperoxidase 유전자 발현에 대한 All-Trans Retinoic Acid의 영향

임규,곽상태,황병두 충남대학교 의과대학 지역사회의학연구소 1994 충남의대잡지 Vol.21 No.1

Effects of all-trans retinoic acid on DNA synthesis and myeloperoxidase (MPO) gene expression have been investigated during the differentiation of human promyelocytic leukemia cell line HL-60. DNA synthesis was decreased at 24hr after exposure with retinoic acid. Conversely control (no adding retinoic acid) gradually increased DNA synthesis. The MPO mRNA was reduced at 24hr in retinoic acid-exposed HL-60 cells. The level of MPO mRNA was also reduced in proportion to the concentration of retinoic acid. MPO mRNA in the retinoic acid-exposed cells was constant after cycloheximide treatment. Retinoic acid-induced repression of MPO gene was no effect by staurosporin treatment. The results suggest that retinoic acid-induced repression of MPO gene is not related to phosphorylation by protein kinase C.

일차배양 Sertoli cell 에서 Testosterone 에 의한 세포성 핵암유전자 발현에 관한 연구

임규,유지헌,김계영,권기량,황병두 ( Kyu Lim,Ji Hun Yoo,Kye Young Kim,Gi Ryang Kweon,Byung Doo Hwang ) 생화학분자생물학회 1993 BMB Reports Vol.26 No.1

The expression of the nuclear protooncogenes have been associated with the cell proliferation and differentiation into changes in nuclear function. To evaluate the possibility that the nuclear protooncogenes play roles testosterone-dependent gene regulation, the effects of testosterone on the expression of the nuclear protooncogenes have been investigated in primary Sertoli cell cultures. c-Myc, c-jun and c-fos were expressed in steady state of primary Sertoli cells but c-myb was not detected. Testosterone increased c-myc and c-jun mRNA with maximal stimulation reached in 16 h. The induction of c-myc was dependent on the concentrations of testosterone. The testsoterone induced c-myc mRNA level was also increased in the cells treated with cycloheximide, but reduced by actinomycin D pretreatment. Even in the absence of testosterone, c-myc mRNA was clearly detectable in Sertoli cells from 8-days-old rats, but hardly detectable in cells from 14 and 28 days of age. Testosterone stimulated c-myc expression in the Sertoli cells from 8 and 14-days-old rats. These results suggest that transient expression of c-myc may play some roles to an intermediate step required for testosterone-dependent regulation of other genes expression of Sertoli cells.

졸겔법으로 합성된 Mal₂O₄:Eu²⁺(M=Sr,Ba,Ca)의 특성

임규,김용현,이형직,김세기,이형복,Lim, Kyu,Kim, Young-Hyun,Lee, Hyung-Jik,Kim, Sei-Ki,Lee, Hyung-Bock 한국세라믹학회 2009 한국세라믹학회지 Vol.46 No.6

Phosphors of $Eu^{2+}$ doped alkaline earth aluminates MA$l_2O_4:Eu^{2+}$ (M=Sr, Ba, Ca) have been prepared by sol-gel process and their characterization of photoluminescence and photocurrent properties have been performed. The phosphors prepared by sol-gel process, due to its advantage of better homogeneity and low synthetic temperature, was synthesized as single phases at lower temperature than the solid-state process; $800{\sim}1000{^{\circ}C}$ for 6 h under mild reduction atmosphere (Ar- 3% $H_2$). It was confirmed that SrA$l_2O_4:Eu^{2+}$ composition revealed the most excellent properties in the brightness and photocurrent.

REGULATION OF TESTIS - SPECIFIC HISTONE GENES DURING SPERMATOGENESIS

임규,chi Bom Chae 생화학분자생물학회 1991 생화학분자생물학회 춘계학술발표논문집 Vol.- No.-

졸겔법으로 합성된 MAl2O4:Eu2+(M=Sr,Ba,Ca)의 특성

임규,김용현,김세기,이형복,이형직 한국세라믹학회 2009 한국세라믹학회지 Vol.46 No.6

Phosphors of Eu2+ doped alkaline earth aluminates MAl2O4:Eu2+ (M=Sr, Ba, Ca) have been prepared by sol-gel process and their characterization of photoluminescence and photocurrent properties have been performed. The phosphors prepared by sol-gel process, due to its advantage of better homogeneity and low synthetic temperature, was synthesized as single phases at lower temperature than the solid-state process; 800~1000 oC for 6 h under mild reduction atmosphere (Ar- 3% H2). It was confirmed that SrAl2O4:Eu2+ composition revealed the most excellent properties in the brightness and photocurrent.

Sertoli 세포의 생리기능 및 호르몬에 의한 조절

임규,박청,윤경아,윤은진,박종일,박승길,황병두 대한내분비학회 2003 Endocrinology and metabolism Vol.18 No.2

骨格筋 S-Adenosyl-L-Methionine : Protein-Carboxyl O-Methyltransferase의 精製 및 特性 Protein-Carboxyl O-Methyltransferase from Bovine Skeletal Muscle

林圭,洪承元,郭相太,黃炳斗 충남대학교 의과대학 지역사회의학연구소 1987 충남의대잡지 Vol.14 No.2

S-Adenosyl-L-methionine : protein-carboxyl 0-methyltransferase(prorein methylase II) has been purified from bovine skeletal muscle approximately 6,600-fold with a 7% yield by combination of ammonium sulfate precipitate, Sephadex G-75 chromatography, S-adenosyl-L-homocysteine-Sepharose 4B chromatography and hydroxyapatite chromatography, and then its characterizations and possible roles were investigated. The enzyme showed a optimum pH around 6.0. It was heat labile, inactivated by heat-treatment at 60℃ for 6 minutes, and when stored at -20℃ in the presence 10% glycerol, its activity has almost stabilized in 6 months. Copper ion(Cu^2+)and zinc ion(Zn^2+)were a potent inhibitors, the activity being completely inhibited at 0.25 mM and 1 mM respectively, The inhibition of copper and zinc ions were recovered 94%, 75% of its activity by adding 4 mM of EDTA, respectively. The apparent Km value for S-adenosyl--L-methionine was 2.86 x 10^-6 M. Kinetic analysis of enzyme in the presence of 30 uM copper ion showed that the nature of the inhibition to the enzyme was noncompetitive considering that Km was constant and Vmax was decreased. Histone IIA, immunoglobulin A and trypsin inhibitor were relatively good substrates for the enzyme. The enzyme activity was completely inhibited by 0.5 mM p-hydroxymercuribenzoic acid, but all of the activity was recovered in adding 10 mM mercaptoethanol, and the molecular weight of the enzyme was 24,000. Sarcolemma and sarcoplasmic reticulum of the skeletal muscle were good substrates for this enzyme and ^3H-CH_3 group was incorporated to 0.63 and 0.96 pmoles/mg protein/min, respectively. The carboxylmethylation of the sarcolemma of the skeletal muscle occurred in the 5 protein fractions(M. W. : 45,000, 22,000, below 14,400etc.) and that of sarcoplasmic reticulum occurred in the 7 protein fractions (M. W. : 100,000, 45,000, 22,000, below 14, 400 etc) by SDS/polyacrylamide gel electrophoresis. Among these molecular weight 22,000 was strongly methylated. These results discuss that protein methylase Il may play some roles in regulating physiological function of sarcolemma and sarcoplasmic reticulum of skeletal muscle.