• Title/Summary/Keyword: E6

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Intratypic Variants of HPV-16 E6jE7 Oncogene Isolated from Sexually High-Risk Women in Busan. (부산지역 유흥업소 종사여성으로부터 분리된 HPV16형의 발암유전자(E6/E7) 돌연변이 유형 분석)

  • Min, Sang-Kee;Kim, Sung-Soon;Choi, Byeong-Sun;Jang, Dai-Ho;Lee, Mee-Ok;Choi, Seung-Hwa;Kim, Nam-Ho;Park, Yon-Koung;Jeong, Yeong-A;Kim, Seong-Joon;Bin, Jae-Hun;Park, Ho-Kuk
    • Journal of Life Science
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    • v.19 no.6
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    • pp.765-769
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    • 2009
  • Recent studies have reported that the distribution of HPV-16 sequence variation differs geographically, and more specifically that HPV-16 E6/E7 intratypic variants might carry a high risk for development of ICC (invasive cervical cancer) and CIN (cervical intraepithelial neoplasia) in a given population. To investigate the genetic diversities of HPV-16 E6/E7 oncogene by region, we collected nineteen HPV-16 isolates from sexually high-risk women in Busan, and analyzed the HPV-16 E6/E7 coding regions (nt 34 to 880) with HPV-16 E6/E7 specific PCR amplification. At the nucleotide levet eleven variants of the E6 genes and nine variants of the E7 genes were identified as follows: E6 T178G (n=l1), E6 T178A (n=l), E6 T350G (n=3), E6 A442C (n=2), E6 AI04T, E6 All1G, E6 C116T, E6 G145T, E6 T183G, E6 C335T, E6 G522C and E7 A647G (n=12), E7 A645C, E7 A777C, E7 G663A, E7 T732C, E7 T760C, E7 A775T, E7 T789C and E7 T795G, respectively. At the amino acid levet the isolated HPV-16 E6 and E7 genes showed eleven E6 variants: E6 D25E (n=12), E6 L83V (n=4), E6 E113D (n=2), E6 MIL, E6 Q3R, E6 P5S, E6 Q14H, E6 D25N, E6 127R, E6 H78Y, E6 C140S and three E7 variants: N29S (n=12), L28F, T72S. HPV16 E6 L83V, the dominant variant in the Caucasian population, showed relatively low frequencies in our study population. We elucidated that the dominant HPV-16 E6/E7 variants were HPV-16 E6 D25E (63.2%) and HPV-16 E7 N29S (63.2%), which were phylogenetically included in Asian lineage. Further study is needed to evaluate the risk of cervical cancer related HPV-16 E6/E7 intratypic variants in the Korean population.

Activities of E6 Protein of Human Papillomavirus 16 Asian Variant on miR-21 Up-regulation and Expression of Human Immune Response Genes

  • Chopjitt, Peechanika;Pientong, Chamsai;Bumrungthai, Sureewan;Kongyingyoes, Bunkerd;Ekalaksananan, Tipaya
    • Asian Pacific Journal of Cancer Prevention
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    • v.16 no.9
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    • pp.3961-3968
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    • 2015
  • Background: Variants of human papillomavirus (HPV) show more oncogenicity than do prototypes. The HPV16 Asian variant (HPV16As) plays a major role in cervical cancer of Asian populations. Some amino acid changes in the E6 protein of HPV16 variants affect E6 functions such as p53 interaction and host immune surveillance. This study aimed to investigate activities of HPV16As E6 protein on modulation of expression of miRNA-21 as well as interferon regulatory factors (IRFs) 1, 3, 7 and c-fos. Materials and Methods: Vectors expressing E6 protein of HPV16As (E6D25E) or HPV16 prototype (E6Pro) were constructed and transfected into C33A cells. HCK1T cells expressing E6D25E or E6Pro were established by transducing retrovirus-containing E6D25E or 16E6Pro. The E6AP-binding activity of E6 and proliferation of the transfected C33A cells were determined. MiR-21 and mRNA of interesting genes were detected in the transfected C33A cells and/or the HCK1T cells, with or without treatment by culture medium from HeLa cells (HeLa-CM). Results: E6D25E showed binding activity with E6AP similar to that of E6Pro. Interestingly, E6D25E showed a higher activity of miR-21 induction than did E6Pro in C33A cells expressing E6 protein. This result was similar to the HCK1T cells expressing E6 protein, with HeLa-CM treatment. The miR-21 up-regulation significantly corresponded to its target expression. Different levels of expression of IRFs were also observed in the HCK1T cells expressing E6 protein. Interestingly, when treated with HeLa-CM, IRFs 1, 3 and 7 as well as c-fos were significantly suppressed in the HCK1T cells expressing E6D25E, whereas those in the HCK1T cells expressing E6Pro were induced. A similar situation was seen for IFN-${\alpha}$ and IFN-${\beta}$. Conclusions: E6D25E of the HPV16As variant differed from the E6 prototype in its activities on epigenetic modulation and immune surveillance and this might be a key factor for the important role of this variant in cervical cancer progression.

Studies on Esterase of Pieris rapae L. II. Biochemical Properties and Immunological Studies (배추흰나비(Pieris rapae L.)의 esterase에 관한 연구 II. 생화학적 특성 및 면역학적 연구)

  • 박철호;김학열;여성문
    • The Korean Journal of Zoology
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    • v.33 no.3
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    • pp.337-345
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    • 1990
  • The properties of three esterases (E2, E6 and E11) which were previously purified from Pieris rapae L. were determined and physiological role of E6 was inferred using immunological methods. Based on inhibitor study, all of the purified esterases were found to be carboxylesterases (EC 3.1.1.1). The Km values for E2, E6 and E11 were determined to be 6.89 X 10-$^4$M. 3.19 $\times$ l0-$^4$M and 3.69 X 10-$^4$M, respectively. The molecular weights of E2, E6 and E11 were estimated to be 42 KD, 81 KD and 174 KD, respectively. The isoelectric points of E2, E6 and E11 were estimated to be pH 5.54, pH 5.89 and pH 6.50, respectively. The concentration of E6 during development was highest at the late 5th instar larval stage and that according to organs at the same stage was highest in midgut. These results suggest that E6 might be a hydrolase involved in the digestion of dietary lipids.

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A New Synthesis of 6,10,14,18-Tetramethyl-5,9,13,17 (E,E,E)-nonadecatetraen-2-one

  • Park, Oee-Sook;Ahn, Su-Kyung
    • Archives of Pharmacal Research
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    • v.16 no.4
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    • pp.265-270
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    • 1993
  • 6, 10, 14, 18-Tetramethyl-5, 9, 131, 17(E, E, E)-nonadecatetraen-2-one was synthesized from geraniol or 6-methyl-5(E)-hepten-2-one in 6 steps, repectively. The key step in both syntheses, was the condensation of phenyl sulfone compound and allylic chloride.

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Identification and Characterization of a Conserved Baculoviral Structural Protein ODVP-6E/ODV-E56 from Choristoneura fumiferana Granulovirus

  • Rashidan, Kianoush Khajeh;Nassoury, Nasha;Giannopoulos, Paresa N.;Guertin, Claude
    • BMB Reports
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    • v.35 no.6
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    • pp.595-603
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    • 2002
  • A gene that encodes a homologue to baculoviral ODVP-6E/ODV-E56, a baculoviral envelope-associated viral structural protein, has been identified and sequenced on the genome of Choristoneura fumiferana granulovirus (ChfuGV). The ChfuGV odvp-6e/odv-e56 gene was located on an 11-kb BamHI subgenomic fragment using different sets of degenerated primers, which were designed using the results of the protein sequencing of a major 39 kDa structural protein that is associated with the occlusion-derived virus (ODV). The gene has a 1062 nucleotide (nt) open-reading frame (ORF) that encodes a protein with 353 amino acids with a predicated molecular mass of 38.5 kDa. The amino acid sequence data that was derived from the nucleotide sequence in ChfuGV was compared to those of other baculoviruses. ChfuGV ODVP-6E/ODV-E56, along with othe baculoviral ODVP-6E/ODV-E56 proteins, all contained two putative transmembrane domains at their C-terminus. Several putative N-and O-glycosylation, N-myristoylation, and phosphorylation sites were detected in the ChfuGV ODVP-6E/ODV-E56 protein. A similar pattern was detected when a hydrophobicity-plots comparison was performed on ChfuGV ODVP-6E/ODV-E56 with other baculoviral homologue proteins. At the nucleotide level, a late promoter motif (GTAAG) was located at -14 nt upstream to the start codon of the GhfuGV odvp-6e/odv-e56 gene. a slight variant of the polyadenylation signal, AATAAT, was detected at the position +10 nt that is downstream from the termination signal. A phylogenetic tree for baculoviral ODVP-6E/ODV-E56 was constructed using a maximum parsimony analysis. The phylogenetic estimation demonstrated that ChfuGV ODVP-6E/ODV-E56 is most closely related to those of Cydia pomonella granulovirus (CpGV) and Plutella xylostella granulovirus (PxGV).

Synthesis of Adenosine and Uridine Analogues Containing Conjugated Diene (공액 이중 결합을 갖는 Adenosine과 Uridine 유사체의 합성)

  • Ro, Bong Oh
    • Journal of the Korean Chemical Society
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    • v.45 no.4
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    • pp.312-317
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    • 2001
  • 7'-Aldehyde-nucleosides analogue (2a, 2c) were synthesized from 6-N-benzoyl-2',3'-O-isopropylideneadenosine and uridine. The condensation of 2 with ethoxycarbonylmethylene Wittig reagent produced the adenosine and uridine analogues containing extended conjugated diene and ethoxycarbonyl group, ethyl-1',5',6',7',8'-pentadeoxy-1'-(adenin-9-yl)-$\beta$-D-ribo-nona-5'(E),7'(E)-dienofuranuronate (4b) and ethyl-1'. 5',6',7',8'-pentadeoxy-1'-(uracil-1-yl)-$\beta$-D-ribo-nona-5'(E),7'-(E)-dienofuranuronate (4c). 9-[8',8'-dibromo-5',6',7',8' -tetradeoxy-$\beta$-D-ribo-octa-5'(E),7'(E)-diene]nucleosides (6b, 6c) were also prepared from 2 with similar method.

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Fluorine Labeling in Biosynthetic Studies (I) : Synthesis of Fluorfarnesols

  • Park, O-Sook
    • Archives of Pharmacal Research
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    • v.9 no.4
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    • pp.237-242
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    • 1986
  • The Synthesis of E, E, E-12-fluorofarnesol and E, Z-6-fluorofarnesol which are key intermediates for the study of biosynthesis of some sesquiterpenes, is described. E, E-Farnesyl acetate is treated with selenium dioxide to give E, E, E-12-hydroxy farnesyl acetate, whih is transformed by DAST into E, E, E-12-hydroxy farnesyl acetate, which is transformed by DAST into E, E, E,-12-fluorofarnesylacetate. The latter compound is hydrolyzed to E, E, E,-12-fluorofarnesol. The reformatsky reaction of 6-methyl-5-hepten-2-one with ethyl bromofluoroacetate affords ethyl 2-fluoro-3-hydroxy-3, 7 dimethyl-6-octanoate. This ester is acetylated and eliminated to give ethyl (Z)-2-fluoro-3, 7-dimethylocta-2, 6-dienoate, which is transformed to allyl bromide via allylic alcohol. The allyl bromide is treated with dianion of methyl acetate to give-keto ester. The $\beta$-keto ester is converted to diethyl phosphoryloxy compound. The conjugate addition of lithium dimethylcuprate to the latter compound gives fluoro ester, which is treated with DIBAL to afford E, Z-6-fluorofarnesol.

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Transcription, Translation, and Immunolocalization of ODVP-6E/ODV-E56 and p74 Proteins: Two Highly Conserved ODV-associated Envelope Proteins of Choristoneura fumiferana Granulovirus

  • Rashidan, Kianoush Khajeh;Nassoury, Nasha;Giannopoulos, Paresa N.;Guertin, Claude
    • BMB Reports
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    • v.38 no.1
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    • pp.65-70
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    • 2005
  • Choristoneura fumiferana granulovirus (ChfuGV) infection results two types of enveloped virions: Occlusion-derived virus (ODV) and budded virus (BV). Structural proteins ODVP-6E/ODV-E56 and p74 are two major conserved ODV-associated proteins that may be involved in the initiation of viral infection cycle in susceptible host insect larvae. This study presents the characterization of ChfuGV odvp-6e/odv-e56 and p74 transcription and translation as well as immunolocalization of these proteins in the occluded ChfuGV virion. Our results revealed that the transcription of odvp-6e/odv-e56 and p74 genes, both, start at 24 hours post infection (h p.i.). Using monospecific polyclonal antibodies made against ODVP-6E/ODV-E56 and p74 we demonstrated that these proteins are both expressed late in infection (24 h p.i.). Immunogold labeling using antisera against ODVP-6E/ODV-E56 and p74 proteins demonstrated that ODVP-6E/ODV-E56 and p74 proteins are both associated with the ODV envelop of ChfuGV.

Establishment of Transgenic Mouse with the E6 and E7 Genes of Human Papillomavirus Type 16 (인간 Papillomavirus의 E6, E7 유전자를 이용한 Transgenic Mouse의 확립)

  • Hwang, Yong-Il;Lee, Seung-Cheol;Kim, Hyun-Su
    • The Journal of Korean Society of Virology
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    • v.26 no.1
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    • pp.115-120
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    • 1996
  • Human papillomavirus (HPV), especially type 16 and 18, has been closely associated with carcinomas and uterine cevical cancer, recently. From in vitro assays, E6 and E7 genes of HPV16 are closely linked with transformation of cell lines of rodent fibroplasts. However, the transforming activity of E6 and E7 genes of HPV type 16 in vivo has not been fully elucidated. For explaining this mechanism, we prepared a expression system with the promoter of mouse mammary tumorvirus long terminal repeat and E6E7's open reading frames. This expression system was introduced in rodent cell lines, No. 7, 3Y1 and shown normal transforming abilities. And, we produced transgenic mice with E6, E7 expression system. These transgenic mice were confirmed from Southern blot analysis. One male of them was observed enlargement of the testis after 5 months postdelivery.

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The Effects of Rhus Extracts on The Cytotoxicity on Cancer Cells and E6 and E7 Oncogenes of Human Papillomavirus Type 16 (옻 추출물의 세포독성 및 자궁 경부암 바이러스 암 유발인자 E6 와 E7의 작용에 미치는 효과)

  • Cho, Young-Sik;Joung, Ok;Cho, Cheong-Weon;Lee, Kyung-Ae;Shim, Jung-Hyun;Kim, Kwang-Soo;Lee, Hong-Soo;Seung, Ki-Seung;Yoon, Do-Young
    • Korean Journal of Food Science and Technology
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    • v.32 no.6
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    • pp.1389-1395
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    • 2000
  • Cervical cancer has been one of the leading causes of female death from cancer worldwide with about 500,000 deaths per year. A strong association between certain human papillomaviruses (HPV types 16 and 18) and cervical cancer has been well known. An extract of natural products, Rhus, has been used to investigate whether this agent has the ability of inhibiting the oncogenes E6 and E7 of HPV type 16. This Rhus inhibited the proliferation of human cervical cancer cell lines (C-33A, SiHa, Caski) and HaCaT keratinocytes in a dose response manner. In vitro binding assay and ELISA showed that Rhus inhibited the in vitro binding of E6 and E6AP which are essential for the binding and degradation of the tumor suppressor p53. In addition, Rhus inhibited the in vitro binding of E7 and Rb which essential tumor suppressor for the control of cell cycle. The level of mRNA for E6 was also decreased by Rhus while that of E7 mRNA was not changed. Our data suggested that Rhus inhibited the oncogenecity of E6 and E7 of HPV 16 type, thus can be used as a putative anti-HPV agent for the treatment of cervical carcinomas by HPV.

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