• Title/Summary/Keyword: E3 region

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Genetic Variation in the Immunoregulatory Gene of Adenovirus Type 3 (3형 아데노바이러스의 면역조절 유전자 다양성)

  • Choi, Eun Hwa;Kim, Hee Sup;Lee, Hoan Jong
    • Pediatric Infection and Vaccine
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    • v.16 no.2
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    • pp.199-204
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    • 2009
  • Purpose : Various proteins encoded in the early region 3 (E3) of adenoviruses protect cells from being killed by cytotoxic T cells and death-inducing cytokines. We sought to find out whether the genetic heterogeneity of the E3 gene might contribute to the molecular diversity of adenoviruses. Methods : Sequences in the E3 region were analyzed for 14 adenovirus type 3 (Ad3) strains that were isolated from children with lower respiratory tract infections in the Seoul National University Children's Hospital during the period 1991-2000. Full-length adenoviral DNA was purified from the infected A549 cell lysates using a modified Hirt procedure. Results : There was 98% homology between 14 Korean Ad3 strains with a reference strain (M15952). Homology within the Korean Ad3 strains was 98.7%. Variation was found in the region of transcripts 20.1 kDa, 20.6 kDa, truncated 7.7 kDa, 10.3 kDa, 14.9 kDa, and 15.3 kDa. In particular, all 14 Korean strains showed a missense single point mutation at the start codon of the truncated 7.7 kDa. In addition, a deletion was found in the truncated 7.7 kDa region by 58 base pairs in 10 strains and 94 base pairs in 4 strains. Variations in amino acids were observed in the receptor internalization and degradation complex (10.3 kDa/14.9 kDa) which stimulates the clearance from the cell surface and subsequent degradation of the receptors for the Fas ligand and TRAIL, while no variations were observed in another immunoregulatory transcript, 19 kDa. Conclusion : Sequence analysis of the immunoregulatory region of adenovirus E3 shows that genetic heterogeneities are related to genome type patterns.

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Association of a p53 Codon 72 Gene Polymorphism with Environmental Factors and Risk of Lung Cancer: a Case Control Study in Mizoram and Manipur, a High Incidence Region in North East India

  • Saikia, Bhaskar Jyoti;Das, Mandakini;Sharma, Santanu Kumar;Sekhon, Gaganpreet Singh;Zomawia, Eric;Singh, Yanglem Mohen;Mahanta, Jagadish;Phukan, Rup Kumar
    • Asian Pacific Journal of Cancer Prevention
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    • v.15 no.24
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    • pp.10653-10658
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    • 2015
  • Background: A very high incidence of lung cancer is observed in Mizoram and Manipur, North East India. We conducted a population based case control study to establish associations of p53 codon 72 polymorphisms and interactions with environmental factors for this high incidence. Material and Methods: A total of 272 lung cancer cases and 544 controls matched for age (${\pm}5years$), sex and ethnicity were collected and p53 codon 72 polymorphism genotypes were analyzed using a polymerase chain based restriction fragment length polymorphism assay. We used conditional multiple logistic regression analysis to calculate adjusted odds ratios and 95% confidence intervals after adjusting for confounding factors. Results: p53 Pro/Pro genotype was significantly associated with increased risk of lung cancer in the study population (adjusted OR=2.14, CI=1.35-3.38, p=0.001). Interactions of the p53 Pro/Pro genotype with exposure to wood smoke (adjusted OR=3.60, CI=1.85-6.98, p<0.001) and cooking oil fumes (adjusted OR=3.27, CI=1.55-6.87, p=0.002), betel quid chewing (adjusted OR=3.85, CI=1.96-7.55, p<0.001), tobacco smoking (adjusted OR=4.42, CI=2.27-8.63, p<0.001) and alcohol consumption (adjusted OR=3.31, CI=1.10-10.03, p=0.034) were significant regarding the increased risk of lung cancer in the study population. Conclusions: The present study provided preliminary evidence that a p53 codon 72 polymorphism may effect lung cancer risk in the study population, interacting synergistically with environmental factors.

Effect of deletion mutants in the regulatory region of transcriptional regulation of glpD and glpE genes (glpD와 glpE 유전자의 조절영역 결손변이주가 전사조절에 미치는 영향)

  • 정희태;최용악;정수열
    • Journal of Life Science
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    • v.5 no.4
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    • pp.162-169
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    • 1995
  • The glpD genes encoding gly-3-p dehydrogenase is essential for the aerobic growth of E. coli on glycerol or gly-3-p. The glpE gene, the function of which is unknownm is transcribed divergently with respect to glpD gene. Expression of the adjacent but divergently transcribed glpD the glpE genes is positively regulated by the cAMP-CRP complex. In this study, for a precise investigation of the functional elements in the regulatory region for transcription activation by cAMP-CRP, deletion mutation have been introducted into the regulatory region. The effect of the deletion mutant on transcriptional regulation was tested in vivo by $\beta$-galctosidase activity. Deletion mutants in the regulatory region of glpD demonstrated that the presence of the CRP-binding site resulted in an sixfold increase in promoter activity. And also deletion mutants of glpE gene demonstrated that the presence of the CRP-binding site resulted in an eightfold increase in promoter activity. Insertion of 22 bp oligomer in the deletion mutants has shown that the CRP binding site is need for maximal expression of glpD and glpE genes. glpD and glpE gene, cAMP-CRP complex, deletion mutant, transcriptional regulation.

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Activity of Human Dihydrolipoamide Dehydrogenase Is Reduced by Mutation at Threonine-44 of FAD-binding Region to Valine

  • Kim, Hak-Jung
    • BMB Reports
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    • v.35 no.4
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    • pp.437-441
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    • 2002
  • Dihydrolipoamide dehydrogenase (E3) is a member of the pyridine nucleotide-disulfide oxidoreductase family. Thr residues are highly conserved. They are at the active site disulfide-bond regions of most E3s and other oxidoreductases,. The crystal structure of Azotobacter vinelandii E3 suggests that the hydroxyl group of Thr that are involved in the FAD binding interact with the adenosine phosphate of FAD. However, several prokaryotic E3s have Val instead of Thr. To investigate the meaning and importance of the Thr conservation in many E3s, the corresponding residue, Thr-44, in human E3 was substituted to Val by site-directed mutagenesis. The mutant’s E3 activity showed about a 2.2-fold decrease. Its UV-visible and fluorescence spectra indicated that the mutant might have a slightly different microenvironment at the FAD-binding region.

Interaction of Tobacco Smoking and Chewing with Angiotensin Converting Enzyme (Insertion/Deletion) Gene Polymorphisms and Risk of Lung Cancer in a High Risk Area from Northeast India

  • Phukan, Rup Kumar;Borah, Prasanta Kumar;Saikia, Bhaskar Jyoti;Das, Mandakini;Sekhon, Gaganpreet Singh;Mahanta, Jagadish
    • Asian Pacific Journal of Cancer Prevention
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    • v.15 no.24
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    • pp.10691-10695
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    • 2015
  • Background: Association of angiotensin converting enzyme (ACE) gene polymorphisms with lung cancer susceptibility remains uncertain and varies with ethnicity. Northeast India represents a geographically, culturally, and ethnically isolated population. The area reports an especially high rate of tobacco usage in a variety of ways of consumption, compared with the rest of the Indian population. Materials and Methods: We conducted a population based case control study in two major high risk region for lung cancer from Northeast India. A total of 151 consecutive lung cancer cases diagnosed histopathologically and equal numbers of controls were recruited with record of relevant sociodemographic information. Blood samples were collected and processed to identify ACE gene polymorphism. Results: Significantly higher (40.4 % vs 29.1%, OR=1.97, CI=1.04-3.72; p=0.037) prevalence of the ACE II genotype was observed among lung cancer cases. Smoking was significantly associated with increased risk of lung cancer (OR=1.70, CI=1.02-2.81; p=0.041). An enhanced risk was also observed for interaction of ACE II genotype with tobacco smoking (OR=4.09, CI=1.51-11.05; p=0.005) and chewing (OR=3.68, CI=1.22-11.13; p=0.021). Conclusions: The present study indicates significant association s of the ACE II genotype with lung cancer in high risk Northeast India.

A Study on the Extraction of E-mail Region in Unconstraint Calling Card Images (무제약 명함 영상에서의 E-mail 영역 검출에 관한 연구)

  • 신상철;정재영
    • Journal of Korea Society of Industrial Information Systems
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    • v.7 no.5
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    • pp.183-189
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    • 2002
  • In this paper, we propose an algorithm to extract the E-mail address in calling card images. Firstly, text regions are separated from background. in the image. To do this, the properties of e-mail addresses and the texture features in the image is used. And then, each text region is explored to find the candidates of e-mail region. Finally, each candidate is divided into characters to find at-symbol(@), that is, e-mail region. The experimental results show hit-ratio over 93.3% for the various kind of calling cards containing different fonts, background images, caricatures.

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Gene Cloning and Nucleotide Sequence of Human Dihydrolipoamide Dehydrogenase-Binding Protein

  • Lee, Jeongmin;Ryou, Chongsuk;Jeon, Bong Kyun;Lee, Poongyeon;Woo, Hee-Jong;Kwon, Moosik
    • Asian-Australasian Journal of Animal Sciences
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    • v.15 no.3
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    • pp.421-426
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    • 2002
  • The pyruvate dehydrogenase complex (PDC), a member of $\alpha$-keto acid dehydrogenase complex, catalyzes the oxidative decarboxylation of pyruvate with the formation of $CO_2$, acetyl-CoA, NADH, and $H^+$. This complex contains multiple copies of three catalytic components including pyruvate dehydrogenase (E1), dihydrolipoamide acetyltransferase (E2), and dihydrolipoamide dehydrogenase (E3). Two regulatory components (E1-kinase and phospho-E1 phosphatase) and functionally less-understood protein (protein X, E3BP) are also involved in the formation of the complex. In this study, we have partially cloned the gene for E3BP in human. Nine putative clones were isolated by human genomic library screening with 1.35 kb fragment of E3BP cDNA as a probe. For investigation of cloned genes, Southern blot analysis and the construction of the restriction map were performed. One of the isolated clones, E3BP741, has a 3 kb-SacI fragment, which contains 200 bp region matched with E3BP cDNA sequences. The matched DNA sequence encodes the carboxyl-terminal portion of lipoyl-bearing domain and hinge region of human E3BP. Differences between yeast E3BP and mammalian E3BP coupled with the remarkable similarity between mammalian E2 and mammalian E3BP were confirmed from the comparison of the nucleotide sequence and the deduced amino acid sequence in the cloned E3BP. Cloning of human E3BP gene and analysis of the gene structure will facilitate the understanding of the role(s) of E3BP in mammalian PDC.

Mtatioal Analysis of the Role of vir-box in the Expression of the virE Gene

  • Han, Seong-Su;Sim, Woong-Seop
    • Journal of Microbiology
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    • v.37 no.3
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    • pp.175-179
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    • 1999
  • To elucidate the role of vir-box in the expression of the virE gene, the vir-box was modified by site-directed mutagenesis and tested for ${\beta}$-galactosidase activities. A, C, T T, A, C substitutions at -62, -63, and -65 positions, destroying the 5'-region of the vir-box and A T at position -55, destroying the 3'-region of the vir-box respectively, showed only 17% promoter activity. When the vir-box was modified to contain perfect dyad symmetry structure (DSR) by the substitutions T, G A, T at -60 an d-61 positions, ${\beta}$-glactosidase activity increased 302%. These results indicate that the 5' and 3'-region of vir-box as well as the imperfect DSR of the vir-box itself may play a very important role in the regulation of virE gene expression.

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Identification of Responsible Region for the Polymerization of Plasmid pEC-3 (Plamid pEC-3의 중합에 필요한 부위의 동정)

  • Jang, Sung-Key;Lee, Ha-Kyu;Rho, Hyune-Mo
    • Korean Journal of Microbiology
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    • v.22 no.3
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    • pp.183-189
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    • 1984
  • In order to find specific acting site of Rec A protein in plasmic polymerization in E. coli, we randomly deleted various part of pEC-3 (a derivative of pBR322) with SI nuclease treatment. Self-ligated plasmids were introduced into E. coli WA802(Rec $A^+$). A number of colonies were analyzed if they contained monomeric or polymeric plasmids by gel electrophoresis. The plasmid (pEC-43), which was deleted the region of tetracycline gene, revealed only monomeric form in Rec $A^+$ E. coli. When two plasmids, pEC-3 and pEC-43, were co-transformed in the same E. coli, the original pEC-3 showed polymerization but pEC-43 revealed monomeric form only. These results suggest that Rec A protein requires the specific site for polymerization.

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Promoter Methylation of MGMT Gene in Serum of Patients with Esophageal Squamous Cell Carcinoma in North East India

  • Das, Mandakini;Sharma, Santanu Kumar;Sekhon, Gaganpreet Singh;Saikia, Bhaskar Jyoti;Mahanta, Jagadish;Phukan, Rup Kumar
    • Asian Pacific Journal of Cancer Prevention
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    • v.15 no.22
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    • pp.9955-9960
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    • 2014
  • Background: Promoter hypermethylation is a common event in human cancer. O6-methylguanine-DNA methyltransferase (MGMT) is a gene involved in DNA repair, which is methylated in a variety of cancers. We aimed to explore the methylation status of MGMT gene among the North Eastern population where esophageal cancer incidence and exposure to carcinogens like nitrosamines is high. Materials and Methods: A total of 100 newly diagnosed esophageal cancer cases along with equal number of age, sex and ethnicity matched controls were included in this study. Methylation specific PCR was used to determine the MGMT methylation status in serum samples. Results: Aberrant promoter methylation of the MGMT gene was detected in 70% of esophageal cancer cases. Hypermethylation of MGMT gene was found to be influenced by environmental factors like betel quid and tobacco which contain potent carcinogens like nitrosamines. Tobacco chewing and tobacco smoking habit synergistically with MGMT methylation elevated the risk for esophageal cancer development [adjusted OR=5.02, 95% CI=1.35-18.74; p=0.010 for tobacco chewing and Adjusted OR=3.00, 95% CI=1.22-7.36; p=0.014 for tobacco smoking]. Conclusions: Results suggest that the DNA hypermethylation of MGMT is an important mechanism for MGMT gene silencing resulting in esophageal cancer development and is influenced by the environmental factors. Thus MGMT hypermethylation can be used as a biomarker for esophageal cancer in high incidence region of North East India.