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Identification of Protein Phosphatase 4 Inhibitory Protein That Plays an Indispensable Role in DNA Damage Response

  • Park, Jaehong;Lee, Jihye;Lee, Dong-Hyun
    • Molecules and Cells
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    • v.42 no.7
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    • pp.546-556
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    • 2019
  • Protein phosphatase 4 (PP4) is a crucial protein complex that plays an important role in DNA damage response (DDR), including DNA repair, cell cycle arrest and apoptosis. Despite the significance of PP4, the mechanism by which PP4 is regulated remains to be elucidated. Here, we identified a novel PP4 inhibitor, protein phosphatase 4 inhibitory protein (PP4IP) and elucidated its cellular functions. PP4IP-knockout cells were generated using the CRISPR/Cas9 system, and the phosphorylation status of PP4 substrates (H2AX, KAP1, and RPA2) was analyzed. Then we investigated that how PP4IP affects the cellular functions of PP4 by immunoprecipitation, immunofluorescence, and DNA double-strand break (DSB) repair assays. PP4IP interacts with PP4 complex, which is affected by DNA damage and cell cycle progression and decreases the dephosphorylational activity of PP4. Both overexpression and depletion of PP4IP impairs DSB repairs and sensitizes cells to genotoxic stress, suggesting timely inhibition of PP4 to be indispensable for cells in responding to DNA damage. Our results identify a novel inhibitor of PP4 that inhibits PP4-mediated cellular functions and establish the physiological importance of this regulation. In addition, PP4IP might be developed as potential therapeutic reagents for targeting tumors particularly with high level of PP4C expression.

The Effects of Gunyuljejo-tang on the CCl4-induced Liver Damage in Rat (건율제조탕이 CCl4로 유발(誘發)된 간손상(肝損傷) 백서(白鼠)에 미치는 영향(影響))

  • Kim, Jung-Yul;Kim, Hyuk;Yang, Sang-Mook;Kim, Dal-Rae;Jeon, Jong-Weon
    • Journal of Sasang Constitutional Medicine
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    • v.16 no.3
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    • pp.96-107
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    • 2004
  • 1. Objectives This study was carried out to investigate the effects of Gunyuljejo-tang on the $CCl_4$-induced Liver Damage in Rats. 2. Methods Sprague-Dawley rats were devided into 5 experimental groups : Normal, $NS+CCl_4$(Solid extract of $CCl_4$ injection group after Normal Saline feed), $GYJJT+CCl_4$(Solid extract of $CCl_4$ injection group after Gunyuljejo-tang feed), $CCl_4+NS$(Normal Saline feed group after $CCl_4$ injection), $CCl_4+GYJJT$(Solid extract of Gunyuljejo-tang feed group after $CCl_4$ injection). Biochemical assays for serum enzyme activities such as AST, ALT, ALP, BUN, Creatinine, Uric Acid, Total Protein, Albumin, Total Cholesterol, Triglyceride, Glucose, and mRNA Revelation of Cytochrome p450 and activities such as LPO, GSH, GST, Glutathione Reductase, Glutathione Peroxidase, SOD, Catalase, Hydroxyproline, and ${\beta}$-Glucuronidase were performed. 3. Results (1) $GYJJT+CCl_4$ showed lower revelation of Cytochrome p450. (2) $GYJJT+CCl_4$ showed higher GSH activity than $NS+CCl_4$, $CCl_4+GYJJT$ showed higher GSH activity than $CCl_4+NS$ injection significantly. (3) $GYJJT+CCl_4$ showed higher GST activity than $NS+CCl_4$. $CCl_4+GYJJT$ showed higher GST activity than $CCl_4+NS$ significantly. (4) $GYJJT+CCl_4$ showed higher Glutathione Peroxidase activity than $NS+CCl_4$, $CCl_4+GYJJT$ showed higher Glutathione Peroxidase activity than $CCl_4+NS$ significantly. (5) $CCl_4+GYJJT$ showed higher SOD activity than $CCl_4+NS$ significantly. (6) $CCl_4+GYJJT$ showed higher Catalase activity than $CCl_4+NS$ significantly. (7) $GYJJT+CCl_4$ showed lower Hydroxyproline than $NS+CCl_4$ significantly, $CCl_4+GYJJT$ showed higher Hydroxyproline than $CCl_4+NS$ significantly. (8) $GYJJT+CCl_4$ showed higher ${\beta}$-Glucuronidase activity than $NS+CCl_4$, $CCl_4+GYJJT$ showed higher ${\beta}$-Glucuronidase activity than $CCl_4+NS$ significantly. 4. Conclusions Gunyuljejo-tang has the recovering effects on the $CCl_4$-induced Liver Damage significantly.

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4,4'-Diaminodiphenylmethane Migration from Nylon Kitchen Utensils using LC-MS/MS (LC-MS/MS를 이용한 나일론수지제 주방기구 중 4,4'-Diaminodiphenylmethane 이행량 실태조사)

  • Eom, Mi-Ok;Yoon, Hae-Jung;Choi, Hyun-Cheol;Jeon, Dae-Hoon;Kim, Hyung-Il;Sung, Jun-Hyun;Park, Na-Young;Lee, Eun-Joon;Lee, Young-Ja
    • Journal of Food Hygiene and Safety
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    • v.24 no.3
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    • pp.211-216
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    • 2009
  • In order to cope with risk issues of overseas on 4,4'-diaminodiphenylmethane (4,4'-MDA) known to used in the production of polyamide resins, we investigated on 4,4'-MDA migration from plastic cooking utensils. 93 samples of plastic cooking utensils were collected from the retail markets in 7 major cities of Korea during March to June, 2007 and 4,4'-MDA levels migrated to food simulants, water, 4% acetic acid, 20% ethanol and n-heptane, on plastic cooking utensils was quantified by liquid chromatography with tandem mass spectrometry (LC-MS/MS). 4,4'-MDA was detected in 14 samples, and the levels of 4,4'-MDA migrated to food simulants were in the range of $0.001{\sim}0.026$ ppm. Among them, the 4,4'-MDA migration amounts of 2 samples were above 0.01 ppm defined by the EU.

Selection of Hypo- and Hyper-tetraploid Seedlings from Abnormal Cotyledons Seedlings Obtained during Crossing of Tetraploid Grapes (Vitis Complexes) (4배체 포도간 교배된 이상자엽실생으로부터 저·고4배체 식물의 선발)

  • Park, Young-Sik;Kim, In-Jong;Park, Sung-Min
    • Horticultural Science & Technology
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    • v.28 no.5
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    • pp.810-817
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    • 2010
  • We observed abnormal morphology of cotyledons occurring in seedlings derived from open-pollinated and cross-pollinated tetraploid grapes and selected aneuploids, especially hypo- and hyper-tetraploid in seedlings with abnormal morphology of cotyledons. Five types of morphologically abnormal cotyledons were observed. In open-pollination of four tetraploid grapes, the frequency of abnormal cotyledons was 1.6% (49 of 3029 seeds). Percentage of aneuploids in the seedlings of abnormal cotyledons was 20.4% (10 of 49 seedlings). Aneuploids in open-pollination consisted of three (4n = 4x-2), four (4n = 4x-1), and three (4n = 4x+1) seedlings. In cross-pollination of tetraploids, the frequency of abnormal cotyledons was 3.4% (59 of 1729 seeds). Percentage of aneuploids in the seedlings with abnormal cotyledons was 22.0% (13 of 59 seedlings). Aneuploids from cross-pollination of tetraploids consisted of two (4n = 4x-2), nine (4n = 4x-1), one (4n = 4x+1), and one (4n = 4x+3) seedlings. According to the results, although the abnormal cotyledon morphology of seedlings obtained from crossing between tetraploid grapes appeared at low rate (2.3%), aneuploid seedlings occurred at high rate (22.0%); therefore, it indicated that this selection strategy might be very efficient in the initial seedling stage.

Synthesis of Novel 1,2-Diazepino[3,4-b]quinoxaline and Pyridazino[3,4-b]quinoxaline Derivatives (새로운 1,2-Diazepino[3,4-b]quinoxaline과 Pyridazino[3,4-b]quinoxaline)

  • Kim, Ho Sik;Jeong, Geuk
    • Journal of the Korean Chemical Society
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    • v.43 no.3
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    • pp.302-306
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    • 1999
  • The 1,3-dipolar cycloaddition reaction of the quinoxaline 4-oxide 10 with 2-chloroacrylonitrile gave the 2,3-dihydro-lH-1,2-diazepino[3,4-blquinoxalines lla, b, respectively, which were converted into the 2,3,4,6-tetrahydro-lH-l,2-diazepino[3,4-b]quinoxaline 12. The reaction of compound lla with selenium dioxide in acetic acid/water resulted in ring transformation to give the 1,4-dihydro-4-oxopyridazino[3,4-blquinoxaline 13.

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Synthesis and Cobalt-Complexes of 1,2,3,4,5,6-Hexaalkyl-1,4-dibora-2-cyclohexene (1,2,3,4,5,6-Hexaalkyl-1,4-dibora-2-cyclohexene의 생성과 착물 생성)

  • Uhm, Jae-Koouk;An, Hee-Won
    • Journal of the Korean Chemical Society
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    • v.41 no.11
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    • pp.586-589
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    • 1997
  • 1,2,3,4,5,6-Hexaalkyl-1,4-dibora-2-cyclohexene was not produced by the general preparation method in our laboratory because of difficulties in the preparation of $B_2Cl_4.$ 2,3-Dietyl-1,4,5,6-tetramethyl-1,4-dibora-2-cyclohexene, 2 was synthesized by the reduction-oxidation reaction of potassium and methyliodide from 1,4-dimethyl-2,3-diethyl-1,4-dibora-2-cyclohexene, 1 as a substrate. Cobalt sandwich-complexes, 6, 7 and 8 were synthesized by the reaction of 2,3-diethyl-1,4,5,6-tetramethyl-1,4-dibora-2-cyclohexene with $({\eta}5-C_5H_5Co(C_2H_4)_2.$.

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Characterization of Lactate Dehydrogenase in Acanthogobius hasta (풀망둑(Acanthogobius hasta) 젖산탈수소효소의 특성)

  • Yum, Jung-Joo
    • Journal of Life Science
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    • v.18 no.2
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    • pp.264-272
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    • 2008
  • The lactate dehydrogenase (EC 1.1.1.27, LDH) isozymes in tissues from Acanthogobius hasta were characterized by biochemical, immunochemical and kinetic methods. The activities of LDH in skeletal muscle and eye tissues were 65.30 and 53.25 units, but LDH activities in heart and liver tissues were very low. LDH/CS (EC 4.1.3.7, citrate synthase) in skeletal muscle was the highest as 22.29. Specific activities of LDH in brain, eye and skeletal muscle were 56.45, 38.04 and 11.0 units/mg, respectively. The LDH isozymes in tissues were separated by polyacrylamide gel electrophoresis after immunoprecipitation with antiserum against $A_4,\;B_4$ eye-specific $C_4$ and liver-specific $C_4$. LDH $AC_4$ isozymes were detected predominantly in skeletal muscle, brain and eye tissues, and $B_4$ isozyme was detected in heart. Anodal eye-specific $C_4$ and cathodal liver-specific $C_4$ were coexpressed in A. hasta. The eye-specific $C_4$ isozyme showed higher activity in eye tissue, but liver-specific $C_4$ isozyme showed lower activity in liver. As a result, one part of molecular structures in $A_4\;and\;C_4,\;A_4\;and\;B_4$, and eye-specific $C_4$ and liver-specific $C_4$ were similar, but in $B_4\;and\;C_4$ were different with each other. Therefore the subunit A may be conservative in evolution, and the evolution of subunit B seems to be faster than that of subunit A. The LDH $A_4$ isozyme of skeletal muscle was purified in the fraction from elution with NAD+ containing buffer of affinity chromatography and eye-specific $C_4$ isozyme was eluted right after $A_4$, so the structure of eye-specific $C_4$ isozyme is similar to $A_4$. And LDH activity remained 35.22-43.47% as a result of the inhibition by pyruvate, the Michaelis-Menten constant values for pyruvate was 0.080-0.098 mM, and Vmax were 153.85 units, 35.09 units in skeletal muscle and eye, respectively. Also the $B_4$ isozyme was the thermo-stablest and $C_4$ was stabler than $A_4$ isozyme. The optimum pH of LDH was 6.5. The results mentioned above indicate that isozymes in tissues showed the properties between LDH $A_4\;and\;B_4$ isozyme as A. hasta was adapted to hypoxic conditions. Also LDH seems to function more effectively under anaerobic condition because LDH in skeletal muscle and eye tissues have high affinity for pyruvate.

Synthesis of Nickel Double-decker Complexes Containing 1,4-Dibora-2,5-cyclohexadiene Derivatives as Ligand (1.4-Dibora-2,5-cyclohexadiene 유도체를 리간드로 갖는 니켈 Double-decker 착물의 합성)

  • Eom, Jae Guk
    • Journal of the Korean Chemical Society
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    • v.46 no.3
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    • pp.213-218
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    • 2002
  • (${\eta}$5-Cyclopentadienyl)(${\eta}$6-1,2,3,4-tetramethyl-1,4-dibora-2,5-cyclohexadiene)Ni (5) was synthe-sizedas a main product by the reaction of cyclopentadienyl(carbonyl)nickel dimer and 1,2,3,4-tetramethyl-1,4-di-bora-2-cyclohexene (1). Di-allyl nickel reacted with 2,3-diethyl-1,4-di-methyl-1,4-dibora-2-cyclohexene (2) at -20$^{\circ}C$ to give bis(${\eta}$6-2,3-diethyl- 1,4-dimethyl-1,4-dibora-2,5-cyclohexadiene)nickel (6) in the yield of 15%. By the reaction of di-allyl nickel and 2,3-dimethyl-1,4-diethoxy-1,4-di- bora-2-cyclohexene (3) at -20$^{\circ}C$ bis[${\eta}$6-2,3-dim-ethyl-1,4-diethoxy-1,4-di- bora-2,5-cyclohexadiene]nickel (7) was obtained in 22% yield. These double-decker complexes were difficult to separate because of their unstabilities and were identified by ESR, NMR, MS and ele-mental analysis, etc.

Expression Study on the Scaffold Gene of CRL4 Complex in Rice (Oryza sativa L.) (벼에 존재하는 CRL4 복합체 scaffold 유전자의 발현 양상에 대한 연구)

  • Bae, Yoowon;Kim, Hani;Kim, Sang-Hoon;Lee, Jae-Hoon
    • Journal of Life Science
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    • v.28 no.10
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    • pp.1132-1139
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    • 2018
  • The stability of diverse cellular proteins in eukaryotes is regulated via ubiquitination. Moreover, E3 ligase plays a crucial role in determining substrate specificity and transfers ubiquitins into the substrates during the ubiquitination process. As a type of multi-subunit E3 ligase, cullin4 (CUL4)-based E3 ligase (CRL4) complex is involved in a variety of cellular processes, such as hormonal and stress responses in plants. In spite of several reports on the versatile roles of CRL4 in various signalings in Arabidopsis, CRL4's function in rice has been poorly known. To learn about CRL4-mediated cellular processes in rice in more detail, OsCUL4 that exhibits the highest homology with Arabidopsis CUL4 was isolated, and its expression patterns in various tissues and in response to plant hormones and abiotic stresses were monitored. Exogenous application of ABA or cytokinin increased the transcript levels of the OsCUL4 gene. Moreover, OsCUL4 was significantly upregulated in response to drought and salt stresses. These findings imply that OsCUL4 may be functionally related to ABA- and/or cytokinin-mediated cellular responses. OsCUL4 directly interacted with OsDDB1, an adaptor protein of CRL4, indicating that OsCUL4 can act as a scaffold protein of CRL4. An expression study on the OsCUL4 gene from this report could be used as a starting point to elucidate cellular responses in which a CRL4-mediated ubiquitination process is involved in rice.

MPEG-4 Contents Authoring using AVI (AVI 데이터를 이용한 MPEG-4 컨텐츠 저작)

  • 이숙영;차경애;김상욱
    • Proceedings of the Korea Multimedia Society Conference
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    • 2002.05d
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    • pp.618-621
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    • 2002
  • MPEG-4 컨텐츠는 비디오, 오디오 파일을 비롯하여 다양한 타입의 미디어로 구성되는 복합 멀티미디어 데이터이다. 본 논문에서는 MPEG-4 컨텐츠 생산을 위해서 저작 환경을 제공하고 이를 MPEG-4 파일로 생성하는 MPEG-4 컨텐츠 저작 도구를 제안한다. 특히 제안하는 MPEG-4 저작 도구는 AVI 형태의 동영상 자료를 입력 받아, MPEG-4 오디오, 비디오 데이터로 변환하고 이를 MPEG-4 장면 저작에 이용할 수 있다. MPEG-4 컨텐츠는 MPEG-4 파일로 구성되며, 장면 구성이 배제된 오디오, 비디오로만 구성된 MPEG-4 파일과 전체 미디어 데이터가 하나의 장면으로 생성된 MPEG-4 파일이 될 수 있다.

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