• Title/Summary/Keyword: Target DNA

Search Result 786, Processing Time 0.028 seconds

Potential role of the histone chaperone, CAF-1, in transcription

  • Kim, Hye-Jin;Seol, Ja-Hwan;Cho, Eun-Jung
    • BMB Reports
    • /
    • v.42 no.4
    • /
    • pp.227-231
    • /
    • 2009
  • The eukaryotic genome forms a chromatin structure that contains repeating nucleosome structures. Nucleosome packaging is regulated by chromatin remodeling factors such as histone chaperones. The Saccharomyces cerevisiae H3/H4 histone chaperones, CAF-1 and Asf1, regulate DNA replication and chromatin assembly. CAF-1 function is largely restricted to non-transcriptional processes in heterochromatin, whereas Asf1 regulates transcription together with another H3/H4 chaperone, HIR. This study examined the role of the yeast H3/H4 histone chaperones, Asf1, HIR, and CAF-1 in chromatin dynamics during transcription. Unexpectedly, CAF-1 was recruited to the actively transcribed region in a similar way to HIR and Asf1. In addition, the three histone chaperones genetically interacted with Set2-dependent H3 K36 methylation. Similar to histone chaperones, Set2 was required for tolerance to excess histone H3 but not to excess H2A, suggesting that CAF-1, Asf1, HIR, and Set2 function in a related pathway and target chromatin during transcription.

Structural Studies of Membrane Protein by Solid-state NMR Spectroscopy (고체상 핵자기공명 분광법을 이용한 막단백질의 구조연구)

  • Kim, Yongae
    • Analytical Science and Technology
    • /
    • v.17 no.5
    • /
    • pp.388-392
    • /
    • 2004
  • Structural studies of membrane proteins, importantly involving interpretation of genomics information, many signaling pathway and major drug target for drug discovery, are having difficulty in characterizing the function using conventional solution nmr spectroscopy and x-ray crystallography because phospholipid bilayers hindered fast tumbling and crystallization. Here, we studied the structure of the pf1 coat protein in oriented phospholipid bilayers by home-built solid-state NMR probe. Bacteriophage pf1 was purified from Paeudomonas Aeruginosa and coat protein of bacteriophage pf1 was isolated from DNA and other proteins.

Molecular cloning and characterization of peroxiredoxin from Toxoplasma gondii

  • Son, Eui-Sun;Song, Kyoung-Ju;Shin, Jong-Chul;Nam, Ho-Woo
    • Parasites, Hosts and Diseases
    • /
    • v.39 no.2
    • /
    • pp.133-141
    • /
    • 2001
  • A cDNA of 1.1 kb comprising the gene encoding the peroxiredoxin of Toxo-plasma gondii(TgPrx) has been cloned. The open reading frame of 591 Up was translated into a protein of 196 amino acids with a molecular mass of 25 kDa. Conserved 2 cysteine domains of Phe-Val-Cys-Pro and Glu-Val-Cys-Pro indicated TgPrx belonged to 2-Cys Prx families. TgPrx showed the highest homology with that of Arabidopsis thaliana by 53.9% followed by Entamoeba histolytica with 39.5% by the amino acid sequence alignment. Polyclonal antibody against recombinant TgPrx detected 25 kDa band in T. gondii without binding to host cell proteins TgPrx was located in the cytoplasm of T. gondii extracellularly or intracellularly by immunofluorescence assay. The expression of TgPrx was increased as early as 30 min after the treatment with artemisinin in the intracellular stage, while no changes in those of host Prx I and TgSOD. This result implies that TgPrx may function as an antioxidant protecting the cell from the attack of reactive oxygen intermediates. It is also suggested that TgPrx is a possible target of chemotherapy.

  • PDF

Isolation of Norfloxacin Resistan Escherichia Coli from the Han River and Characterization of Resistance Mechanism

  • Yoosun Jung;Hunjin Hong;Hyeran Nam;Lee, Yeonhee
    • Journal of Microbiology
    • /
    • v.40 no.1
    • /
    • pp.63-69
    • /
    • 2002
  • A total of twenty-five norfloxacin resistant Escherichia coli were isolated from Joongrang-chun stream, a branch of the Han River in Seoul, Korea from May to July in 2000 and their norfloxacin resistance mechanism was characterized for target site mutation, permeability, and efflux pump. Fourteen iso- lates contained the same three mutations, Ser83→Leu and Asp87→Asn in GyrA and Ser90→ lle in ParC. Six isolates had Ser83→Leu and Asp87→Tyr in GyrA and Ser87→lle in ParC while one isolate had Ser83→Leu and Va1103→Ala in GyrA and Ser80→lle in ParC. Two isolates had mutation(s) in GyrA without any mutation in ParC. Two isolates had Ser80→Arg in ParC instead of the commonly found Ser80→lle. Every norfloxacin resistant isolate had an efflux system but the correlation between the efflux activity and MIC was not observed. The amount of OmpF for norfloxacin permeability decreased in resistant isolates compared to the susceptible strains. When amplified polymorphic DNA (RAPD) and pulse field gel electrophoresis (PFGE) were performed, these isolates showed no similarity to each other or clinical isolates.

An Introduction to Asia Cooperation Programme in Conservation Science(ACPCS, 2005) and the operation result (아시아권 문화재 보존과학 협력과정(ACPCS, 2005)에 대한 소개 및 운영 결과)

  • Jeong, So-Young;Chung, Yong-Jae;Kim, Yong-Han
    • 보존과학연구
    • /
    • s.26
    • /
    • pp.203-211
    • /
    • 2005
  • The National Research Institute of Cultural Heritage(NRICH) conducted the Asia Cooperation Programme in Conservation Science(ACPCS) to facilitate and promote the regional cooperation in the conservation of cultural heritage among Asian countries. This course was promoted to provide an opportunity to study and work together, also to share a knowledge and experience in conservation field. The target countries were from 17 Asian countries, and this year our selection committee selected 2 specialist in cultural heritage conservation field and selected persons had participated in the ACPCS course during three months. NRICH provided the round-trip airline tickets, overseas travel accident insurance and a living allowance to the course participants. And we requested programme announcement to Korean National Commission for UNESCO and Ministry of Foreign Affairs and Trade to progress favorably. This course period was three months from 29 August to 25 November, and the fields were conservation and restoration of tangible cultural heritage such as metal conservation, stone conservation, material analysis, DNA analysis, dating, biological control management of cultural heritage, environmental monitoring and so on.

  • PDF

Cancer Chemoprevention by Dietary Phytochemicals: Rationale and Mechanisms (Dietary Phytochemical을 이용한 화학적 암에방과 그 작용 기전)

  • Surh, Young-Joon;Lee, Jong-Min
    • Environmental Mutagens and Carcinogens
    • /
    • v.18 no.1
    • /
    • pp.1-8
    • /
    • 1998
  • Chemoprevention refers to the use of non-toxic chemical agents to prevent the neoplastic development by inhibiting, delaying, or reversing a multi-stage carcinogenesis. The primary goal of chemoprevention research is to identify or produce effective agents and strategies for clinical trials for applications to normal or high risk human populations. A large number of compounds have been tested for their possible chemopreventive activities, and it is of interest to note that many of them are naturally occurring substances. Thus, a variety of plant and vegetable constituents, particularly those included in our daily diet, have been found to possess substantial protective properties against experimental carcinogenesis. These substances, collectively known as dietary phytochemicals, exert their chemopreventive effects by influencing specific step(s) of multi-stage carcinogenesis: some inhibit metabolic activation or enhance detoxification of carcinogens, others interfere with covalent interactions between ultimate eloctrophilic carcinogens and the target cell DNA and still others may exert anti-promoting or anti-progressing effects. Mechanism-based interventions by use of safe dietary phytochemicals may provide one of the most practical and promising cancer chemopreventive strategies.

  • PDF

Cloning of the Setd1b gene of Mus musculus, a novel histone methyl transferase target in the epigenetic therapy of cancers

  • Morishita, Masayo;Cho, Minju;Ryu, Juhee;Mevius, Damiaan E.H.F.;Di Luccio, Eric
    • Current Research on Agriculture and Life Sciences
    • /
    • v.28
    • /
    • pp.63-68
    • /
    • 2010
  • The epigenetic therapy of cancers is emerging as an effective and valuable approach to both chemotherapy and the chemoprevention of cancer. The utilization of epigenetic targets that include histone methyltransferase (HMTase), Histone deacetylatase, and DNA methyltransferase, are emerging as key therapeutic targets. SET containing proteins such as the HMTase Setd1b has been found significantly amplified in cancerous cells. In order to shed some light on the histone methyl transferase family, we cloned the Setd1b gene from Mus musculus and build a collection of vectors for recombinant protein expression in E.coli that will pave the way for further structural biology studies. We prospect the role of the Setd1b pathway in cancer therapy and detail its unique value for designing novel anti-cancer epigenetic-drugs.

  • PDF

Study of Signaling Pathway on Apoptotic Cell Death Induced by Extract of Ailanthus altissima in Human Jurkat Lymphocytes (저근백피(樗根白皮) 추출물에 의한 급성 림프성 백혈병 Jurkat Lymphocytes의 세포고사 유도 및 신호기전 연구)

  • Lee, Ki Ouk;Kim, Ae Wha;Lim, Kyu Sang;Yun, Young Gab
    • Herbal Formula Science
    • /
    • v.25 no.3
    • /
    • pp.349-362
    • /
    • 2017
  • Objectives : We investigated whether the components of Ailanthus altissima induced apoptotic cell death in Jurkat acute lymphoblastic leukemia (ALL) cells. Methods : Regulation of cell proliferation is a complex process involving the regulated expression and/or modification of discrete gene products, which control transition between different stages of the cell cycle. Results : Upon treatments with Ailanthus altissima, the concentration-dependent inhibitions of cell viability were observed as compared to untreated control group. The capability of Ailanthus altissima to induce apoptosis was associated with proteolytic cleavage of specific target proteins such as poly(ADP-ribose)polymerase (PARP) and beta-catenin proteins suggesting the possible involvement of caspases. Ailanthus altissima also caused apoptosis as measured by cell morphology and DNA fragmentation. Conclusions : These results indicate that the increase of apoptotic cell death by Ailanthus altissima may be due to the inhibition of cell cycle in human Jurkat lymphocytes. Conclusively, these current and further findings will provide novel approaches to understanding and treating major diseases.

Recombination and Expression of eaeA Gene in Enterohemorrhagic Escherichia coli O157:H7

  • Kim, Hong;Kim, Jong-Bae
    • Biomedical Science Letters
    • /
    • v.8 no.3
    • /
    • pp.107-113
    • /
    • 2002
  • Enterohemorrhagic Escherichia coli (EHEC) strains of serotype O157:H7 have been shown to colonize the intestinal epithelial cell by the attaching and effacing (AE) mechanism. The AE lesion is mediated by an intimin, of which production and expression are controlled by a 3-Kb eaeA gene located EHEC chromosomal DNA. If the eaeA gene is mutated, EHEC O157:H7 strains lose capacity of adhesion to intestinal epithelial cells. In this study, a 891 bp of the 3'-end region of a gamma intimin was amplified by polymerase chain reaction (PCR). The PCR product was inserted into pSTBlue-1 cloning vector and transformed into DE3 (BL21) competent cell. After plasmid mini-preparation and restriction enzyme digestion of eaeA/891-pSTBlue-1 vector, target eaeA gene was re-inserted into pET-28a expression vector and was transformed. Then the expression of recombinant eaeA/891 (891 bp) gene was induced by isopropyl-$\beta$-D-thiogalactopyranoside (IPTG). The expression of the 40-KDa recombinant protein was identified in SDS-PAGE and confirmed by immunoblotting using the His.Tag$^{\circledR}$ and T$_{7}$.Tag$^{\circledR}$ monoclonal antibody. This recombinant protein expressed by eaeA gene could be applied in further studies on the mechanisms of E. coli O157:H7 infection and the development of recombinant vaccine.

  • PDF

Identification and Characterization of Protein Arginine Methyltransferase 1 in Acanthamoeba castellanii

  • Moon, Eun-Kyung;Kong, Hyun-Hee;Hong, Yeonchul;Lee, Hae-Ahm;Quan, Fu-Shi
    • Parasites, Hosts and Diseases
    • /
    • v.55 no.2
    • /
    • pp.109-114
    • /
    • 2017
  • Protein arginine methyltransferase (PRMT) is an important epigenetic regulator in eukaryotic cells. During encystation, an essential process for Acanthamoeba survival, the expression of a lot of genes involved in the encystation process has to be regulated in order to be induced or inhibited. However, the regulation mechanism of these genes is yet unknown. In this study, the full-length 1,059 bp cDNA sequence of Acanthamoeba castellanii PRMT1 (AcPRMT1) was cloned for the first time. The AcPRMT1 protein comprised of 352 amino acids with a SAM-dependent methyltransferase PRMT-type domain. The expression level of AcPRMT1 was highly increased during encystation of A. castellanii. The EGFP-AcPRMT1 fusion protein was distributed over the cytoplasm, but it was mainly localized in the nucleus of Acanthamoeba. Knock down of AcPRMT1 by synthetic siRNA with a complementary sequence failed to form mature cysts. These findings suggested that AcPRMT1 plays a critical role in the regulation of encystation of A. castellanii. The target gene of AcPRMT1 regulation and the detailed mechanisms need to be investigated by further studies.