• Title/Summary/Keyword: p21 protein

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Synergistic efficacy of LBH and αB-crystallin through inhibiting transcriptional activities of p53 and p21

  • Deng, Yun;Li, Yongqing;Fan, Xiongwei;Yuan, Wuzhou;Xie, Huaping;Mo, Xiaoyang;Yan, Yan;Zhou, Junmei;Wang, Yuequn;Ye, Xianli;Wan, Yongqi;Wu, Xiushan
    • BMB Reports
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    • v.43 no.6
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    • pp.432-437
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    • 2010
  • LBH is a transcription factor as a candidate gene for CHD associated with partial trisomy 2p syndrome. To identify potential LBH-interacting partners, a yeast two-hybrid screen using LBH as a bait was performed with a human heart cDNA library. One of the clones identified encodes ${\alpha}B$-crystallin. Co-immunoprecipitation and GST pull-down assays showed that LBH interacts with ${\alpha}B$-crystallin, which is further confirmed by mammalian two-hybrid assays. Co-localization analysis showed that in COS-7 cells, ${\alpha}B$-crystallin that is cytoplasmic alone, accumulates partialy in the nucleus when co-transfected with LBH. Transient transfection assays indicated that overexpression of LBH or ${\alpha}B$-crystallin reduced the transcriptional activities of p53 and p21, respectively, Overexpression of both ${\alpha}B$-crystallin and LBH together resulted in a stronger repression of the transcriptional activities of p21 and p53. These results showed that the interaction of LBH and ${\alpha}B$-crystallin may inhibit synergistically the transcriptional regulation of p53 and p21.

Expression Patterns of P21-Activated Kinase 2 by Sterne Spores on Human Macrophages (사람 대식세포에 스턴아포에 처리에 의한 P21-activated kinase 2 단백질의 발현양상)

  • Seo Gwi-Moon;Jung Kwang-Hwa;Kwak Hyun Jung;Kim Seong-Joo;Kim Ji-Chon;Chai Young-Gyu
    • Journal of the Korea Institute of Military Science and Technology
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    • v.7 no.4 s.19
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    • pp.87-90
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    • 2004
  • In order to elucidate the mechanism of infection on human macrophages, we peformed the 2-dimensional electrophoresis and the western blot analysis using the infected human macrophages with the spores of live and inactivated Sterne. We confirmed P21-activated kinase 2 protein which related to cell death(apoptosis) human macrophages at the early stage events. The inhibition of the P21-activated protein kinase 2 protein will be reduced apoptosis on infected human macrophages with Sterne spores.

SAMD4B, a novel SAM-containing protein, inhibits AP-1-, p53- and p21-mediated transcriptional activity

  • Luo, Na;Li, Guan;Li, Yongqing;Fan, Xiongwei;Wang, Yuequn;Ye, Xiangli;Mo, Xiaoyan;Zhou, Junmei;Yuan, Wuzhou;Tan, Ming;Xie, Huaping;Ocorr, Karen;Bodmer, Rolf;Deng, Yun;Wu, Xiushan
    • BMB Reports
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    • v.43 no.5
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    • pp.355-362
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    • 2010
  • The sterile alpha motif (SAM) is a putative protein interaction domain involved in a wide variety of biological processes. Here we report the identification and characterization of a novel gene, SAMD4B, which encodes a putative protein of 694 amino acids with a SAM domain. Northern blot and RT-PCR analysis showed that SAMD4B is widely expressed in human embryonic and adult tissues. Transcriptional activity assays show SAMD4B suppresses transcriptional activity of L8G5-luciferase. Over-expression of SAMD4B in mammalian cells inhibited the transcriptional activities of activator protein-1 (AP-1), p53 and p21, and the inhibitory effects can be relieved by siRNA. Deletion analysis indicates that the SAM domain is the main region for transcriptional suppression. The results suggest that SAMD4B is a widely expressed gene involved in AP-1-, p53- and p21-mediated transcriptional signaling activity.

Seasons affect the phosphorylation of pork sarcoplasmic proteins related to meat quality

  • Zeng, Xianming;Li, Xiao;Li, Chunbao
    • Animal Bioscience
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    • v.35 no.1
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    • pp.96-104
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    • 2022
  • Objective: Sarcoplasmic proteins include proteins that play critical roles in biological processes of living organisms. How seasons influence biological processes and meat quality of postmortem muscles through the regulation of protein phosphorylation remain to be investigated. In this study, the phosphorylation of sarcoplasmic proteins in pork longissimus muscle was investigated in four seasons. Methods: Sarcoplasmic proteins were extracted from 40 pork carcasses (10 for each season) and analyzed through ProQ Diamond staining for phosphorylation labeling and Sypro Ruby staining for total protein labeling. The pH of muscle, contents of glycogen and ATP were measured at 45 min, 3 h, and 9 h postmortem and the water (P2b, P21, and P22) was measured at 3 h and 9 h. Results: A total of 21 bands were detected. Band 8 (heat shock cognate 71 kDa protein; heat shock 70 kDa protein 1B) had higher phosphorylation level in summer than that in other seasons at 45 min postmortem. The phosphorylation levels of 3 Bands were significantly different between fast and normal pH decline groups (p<0.05). The phosphorylation levels of 4 bands showed negative associations with immobilized water (P21) and positive association with free water (P22). Conclusion: The phosphorylation levels of sarcoplasmic proteins involved in energy metabolism and heat stress response at early postmortem time differed depending on the seasons. These proteins include heat shock protein 70, pyruvate kinase, phosphoglucomutase-1, glucose-6-phosphate isomerase, and carbonic anhydrase 3. High temperatures in summer might result in the phosphorylation of those proteins, leading to pH decline and low water holding capacity.

Transcriptional Repression of High-Mobility Group Box 2 by p21 in Radiation-Induced Senescence

  • Kim, Hyun-Kyung;Kang, Mi Ae;Kim, Mi-Sook;Shin, Young-Joo;Chi, Sung-Gil;Jeong, Jae-Hoon
    • Molecules and Cells
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    • v.41 no.4
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    • pp.362-372
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    • 2018
  • High mobility group box 2 (HMGB2) is an abundant, chromatin-associated, non-histone protein involved in transcription, chromatin remodeling, and recombination. Recently, the HMGB2 gene was found to be significantly downregulated during senescence and shown to regulate the expression of senescent-associated secretory proteins. Here, we demonstrate that HMGB2 transcription is repressed by p21 during radiation-induced senescence through the ATM-p53-p21 DNA damage signaling cascade. The loss of p21 abolished the downregulation of HMGB2 caused by ionizing radiation, and the conditional induction of p21 was sufficient to repress the transcription of HMGB2. We also showed that the p21 protein binds to the HMGB2 promoter region, leading to sequestration of RNA polymerase and transcription factors E2F1, Sp1, and p300. In contrast, NF-Y, a CCAAT box-binding protein complex, is required for the expression of HMGB2, but NF-Y binding to the HMGB2 promoter was unaffected by either radiation or p21 induction. A proximity ligation assay results confirmed that the chromosome binding of E2F1 and Sp1 was inhibited by p21 induction. As HMGB2 have been shown to regulate premature senescence by IR, targeting the p21-mediated repression of HMGB2 could be a strategy to overcome the detrimental effects of radiation-induced senescence.

Effects of Arsenic Trioxide on Cell Cycle Related Proteins (Cyclin D1, p21, p27) Expression During Urethane-induced Lung Carcinogenesis in Mice (Urethane으로 유발된 생쥐 폐샘암종 발생과정에서 세포주기 관련인자(Cyclin D1, p21, and p27)에 대한 비소의 효과)

  • Yim Sung-Hyuk;Jeong Ji-Hoon;Gyeon Jong-Man;Park Eon-Sub
    • YAKHAK HOEJI
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    • v.50 no.2
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    • pp.84-92
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    • 2006
  • The present study investigated an effect of arsenic trioxide on the urethane-induced lung carcinogenesis in mice. To understand its carcinogenesis, we examined proliferating cell nuclear antigen (PCNA), apoptotic index as well as cell cycle-related proteins (cyclin D1, p21, and p27). Urethane was injected intraperitoneally in ICR mice, and then they were sacrificed at 5, 15, or 25 weeks following treatment of arsenic trioxide. Arsenic trioxide was given with tap water at a concentration of 1 mg/l (low-dose) and 5mg/1 (high-dose) for 25 weeks. During the carcinogenesis, sequential histological changes from hyperplasia to adenomas, and ultimately to overt carcinomas were noted. The development of hyperplasias, adenomas, and carcinomas in the lung were slightly increased by the treatment of low-dose arsenic trioxide. However, there is no correlation between dose and tumor multiplicity. The administration of low-dose arsenic trioxide, significantly increased the tumor size. The proliferative index observed on 5 weeks after significantly increased. Cyclin D1 and p21 protein, cell cycle related proteins, were more significantly increased in hyperplasia and adenoma in low dose arsenic treated group than urethane alone group. The p27 protein expression did not show any significantly changes with arsenic treated or untreated group. Low dose exposure to arsenic trioxide resulted in increased expression of cyclin D1 and p21 protein. The present results indicate that low-dose treatment of arsenic trioxide, but not high dose of it, partly modulate the cellular proliferation, cyclin D1, and p21 protein expression, and that this effect may contribute to accelerated development of lung adenocarcinomas in urethane-induced mice.

Nutritional Evaluation of Canola Protein Concentrate for Broiler Chickens

  • Thacker, P.A.;Petri, D.
    • Asian-Australasian Journal of Animal Sciences
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    • v.24 no.11
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    • pp.1607-1614
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    • 2011
  • This trial was conducted to determine the effects of including canola protein concentrate in diets fed to broiler chickens on nutrient digestibility and broiler performance (0-21 days). A total of 180, day-old, male broiler chicks weighing an average of 52.8${\pm}$0.6 g were assigned to one of six dietary treatments in a completely randomized design. The control diet was based on corn and soybean meal and contained 15% canola meal. The experimental diets contained 3, 6, 9, 12 or 15% canola protein concentrate added at the expense of canola meal. There were five birds per pen and six replicate pens per treatment. Feed and water were available ad libitum throughout the 21-day experiment. Chromic oxide (0.35%) was added to all diets as a digestibility marker and was fed throughout the experimental period. The digestibility of dry matter, energy and phosphorus increased linearly (p<0.01) with increasing levels of canola protein concentrate. Although nutrient digestibility was higher for birds fed diets containing canola protein concentrate, these improvements did not translate into improvements in broiler performance. Weight gain was unaffected (p = 0.24) by level of canola protein concentrate. Feed intake was significantly increased (p<0.01) with the result that feed conversion tended to be poorer (p = 0.07) for birds fed diets containing canola protein concentrate. Mortality was also unaffected (p = 0.56) by dietary treatment.

Effects of Protein Levels on Growth and Nitrogen Balance in Growing Jindo Dog (단백질 수준이 진도자견의 성장과 질소평형에 미치는 영향)

  • Kang, T.I.;Maeng, W.J.;Kim, M.H.;Lee, Sang-Rak
    • Journal of Animal Science and Technology
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    • v.49 no.5
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    • pp.633-638
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    • 2007
  • This study was conducted to determine the protein requirement level in growing Jindo dog through nitrogen balance experiment. Twelve female dogs aged 18~20 weeks old were allotted one of 3 dietary treatments containing 21, 23 and 25% of crude protein. Average daily gain of dogs fed experimental diets containing 21, 23 and 25% of crude protein were 65.42, 79.58 and 99.17g/d, respectively, and there was a significant difference between 21 and 25% of crude protein treatments(p<0.05). Retained nitrogen were calculated 0.74, 0.96 and 1.31g/kgBW.75/d for dogs fed diets containing 21, 23 and 25% of crude protein, respectively, and were significantly higher(p<0.05) in dogs fed 25% of crude protein diet then those of other dogs. A quadratic regression equation was calculated between nitrogen intake(x) and nitrogen retention(y); y=-2.519x2+12.79x-14.79, and it was found a significantly(p<0.05) higher regression coefficient of 0.782. From the above equation, it was estimated maintenance requirement of crude protein for growing Jindo dog is 11.25g/kg BW.75/d.

Funcyional Studies on Gene 2.5 Protein of Bacteriophage T7 : Protein Interactions of Replicative Proteins (박테리오파아지 T7 의 기능에 관한 연구;복제단백질간의 단백질 상호작용)

  • 김학준;김영태
    • Journal of Life Science
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    • v.6 no.3
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    • pp.185-192
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    • 1996
  • Bacteriophage T7 gene 2.5 protein, a single-stranded DNA binding protein, is required for T7 DNA replication, recombination, and repair. T7 gene 2.5 protein has two distinctive domains, DNA binding and C-terminal domain, directly involved in protein-protein interaction. Gene 2.5 protein participates in the DNA replication of Bacteriophage T7, which makes this protein essential for the T7 growth and DNA replication. What gene 2.5 protein makes important at T7 growth and DNA replication is its binding affinity to single-stranded DNA and the protein-protein important at T7 DNA replication proteins which are essential for the T7 DNA synthesis. We have constructed pGST2.5(WT) encoding the wild-type gene 2.5 protein and pGST2.5$\Delta $21C lacking C-terminal 21 amino acid residues. The purified GST-fusion proteins, GST2.5(WT) and GST2.5(WT)$\Delta$21C, were used for whether the carboxyl-terminal domain participates in the protein-protein interactions or not. GST2.5(WT) and GST2.5$\Delta$21C showed the difference in the protein-protein interaction. GST2.5(WT) interacted with T7 DNA polymerase and gene 4 protein, but GST2.5$\Delta$21C did not interact with either protein. Secondly, GST2.5(WT) interacts with gene 4 proteins (helicase/primase) but not GST2.5$\Delta$21C. these results proved the involvement of the carboxyl-terminal domain of gene 2.5 protein in the protein-protein interaction. We clearly conclude that carboxy-terminal domain of gene 2.5 protein is firmly involved in protein-protein interactions in T7 replication proteins.

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Repression of p21 Expression by Hepatitis B Virus X Protein via a p53-Independent Pathway

  • An, Ji-Yeong;Jang, Gyeong-Rip
    • Proceedings of the Korean Society of Life Science Conference
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    • 2000.12a
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    • pp.39-43
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    • 2000
  • HBV는 인체에 감염하여 간염, 간경변 및 간암을 유발하는 hepadnaviruses의 일종으로써 임상적으로 매우 중요한 바이러스이다. 그러나 이 바이러스에 의한 간암(HCC)의 발생 메커니즘은 아직 불확실하다. 최근에는 HBV의 X 단백질(HBx)이 간암 발생에 중요한 역할을 수행하는 것으로 보고되고 있다. HBx 단백질은 전사 활성인자(transcriptional activator)로써 숙주세포의 유전자발현에 영향을 미치어 세포증식 및 분화에 영향을 줄 수 있다. 본 연구에서는 HBx 단백질이 NIH 3T3 cell의 증식 및 형질전환에 미치는 영향을 조사하였다. HBx 단백질을 발현하는 세포주는 정상세포에 비하여 증식 속도가 2배 정도 빠르며, soft agar assay 결과에 의하면 대조군과 비교하여 더 많은 수의 colony를 형성하였다. 또한, 이들 HBx 발현 세포들은 접촉 저해 능력을 상실하여 HBx가 세포 형질 전환 능력을 가짐을 알수 있다 또한 HBx 발현 세포주에 있어서 p21의 RNA 및 protein수준이 정상세포에 비하여 낮으므로 HBx에 의한 증식 촉진 및 세포 형질 전환이 p21을 매개하여 이루어 짐을 알 수 있었다. HBx에 의한 p21 유전자의 발현 감소는 p21의 전사 수준에서 이루어지며 이는 p53-비의존적 경로에 의하여 이루어졌다.

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