• Title/Summary/Keyword: DNA replication

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Dna2 Helicase/endonuclease Interacts with a Novel Protein YHR122W Protein in Saccharomyces cerevisiae (Saccharomyces cerevisiae에서 Dna2 helicase/endonuclease와 YHR122W 단백질의 상호작용)

  • Lee, Hyun-Sun;Choi, Do-Hee;Kwon, Sung-Hoon;Kim, Na-Yeon;Lee, In-Hwan;Kim, Hyun-Jung;Bae, Sung-Ho
    • Korean Journal of Microbiology
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    • v.42 no.1
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    • pp.1-6
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    • 2006
  • Saccharomyces cerevisiae Dna2 helicase/endonuclease plays an essential role in removing DNA primers during Okazaki fragment processing in eukaryotic DNA replication. Genome-wide scale co-immunoprecipitation experiments predicted that Dna2 interacts with a novel protein YHR122W (1). In this study, we observed that overexpression of YHR122W gene suppressed the temperature-sensitive phenotype of $dna2\Delta405N$ mutation. To investigate direct interaction between these two proteins, a histidine-tagged recombinant YHR122W protein was expressed and purified from E. coli. Physical interaction between the purified YHR122W and Dna2 proteins was detected by enzyme-linked immunosorbent assays. Further more, the complex formation was most efficient at physiological salt concentration, 150 mM NaCl. The genetic and physical interactions between YHR122W and Dna2 shown in this study suggest that the biological functions of these two proteins may be closely related each other.

An Error Detection and Repair on DNA Duplicate Structure (DNA 이중나선구조에서의 오류 검출 및 복구)

  • Kim, Soke-Hwan;Hur, Chang-Wu
    • Journal of the Korea Institute of Information and Communication Engineering
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    • v.15 no.11
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    • pp.2500-2504
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    • 2011
  • Living organisms are composed of cells that can replicate themselves through growth, division packed with tons capacity. On DNA mutations, ie mutations in the offspring's survival and reproduction can be held against you, and packed with tons ambivalence that could benefit. In this study, the DNA double helix is used as a template for replication, we first separated into single strands of the double helix must be opened Combining the double helix portion of the location of errors in the bond provides a way to find and repair.

Isolation and Characterization of New Family Genes of DNA Damage in Fission Yeast

  • Choi, In-Soon
    • Environmental Mutagens and Carcinogens
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    • v.19 no.1
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    • pp.28-33
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    • 1999
  • The SNF2 family includes proteins from a variety of species with roles I cellular processes such as transcriptional regulation, recombination and various types of DNA repair. Several proteins with unknown function are also included in this family. Here, we report the cloning and characterization of hrp 2+ gene (helicase related gene from S. pombe) which was isolated by PCR amplication using the conserved domain of SNF2 motifs within the ERCC6 gene which encodes a protein involved in DNA excision repair. The hrp2+ gene was isolated by screening with yeast S. pombe genomic library. The isolated cloned contained 6.5 kb insert DNA. Southern blot analysis confirmed that S. pombe chromosome contains the same DNA as hrp2+ gene and this gene exists as a single copy in S. pombe genome. The 4.7 kb transcript of mRNA was identified by Northern blot. To examined the transcriptional regulation of hrp2+ gene, DNA damaging agents were treated. These results indicated that the hrp2+ gene may not be directly involved in DNA replication, but may be involved in damage response pathway.

Back to the Ends: Chromosomal DNA (염색체 말단부위)

  • Lee, Mi-Hyung;Suh, Dong-Chul
    • Childhood Kidney Diseases
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    • v.12 no.1
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    • pp.1-10
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    • 2008
  • Nucleic scids transfer the genetic information for serving a central biological purpose. The nucleic acids are polymers of nucleotides and they are mainly ribonucleic acid(RNA) and deoxyribonucleic acid(DNA). The nucleotides are stoichiometrically composed of five-carbon sugars, nitrogeneous bases, and phosphoric acids. The chemistry of nucleic acids and characteristics of different genomes are decribed for further study. Most of DNA genomes tend to be circular including bacterial genomes and eukaryotic mitochondrial DNA. Eukaryotic chromosomes in cells, in contrast, are generally linear. The ends of linear chromosomes are called telomeres. The genomes of different species, such as mammals, plants, invertebrates can be compared with the chromosome ends. The telomeric complex allows cells to distinguish the random DNA breaks and natural chromosomal ends. The very ends of chromosomes cannot be replicated by any ordinary mechanisms. The shortening of telomeric DNA templates in semiconservative replication is occurred with each cell division. The short telomere length is critically related to aging, tumors and dieases.

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Multiple Alternating Immunizations with DNA Vaccine and Replication-incompetent Adenovirus Expressing gB of Pseudorabies Virus Protect Animals Against Lethal Virus Challenge

  • Kim, Seon-Ju;Kim, Hye-Kyung;Han, Young-Woo;Aleyas, Abi G.;George, Junu A.;Yoon, Hyun-A;Yoo, Dong-Jin;Kim, Koan-Hoi;Eo, Seong-Kug
    • Journal of Microbiology and Biotechnology
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    • v.18 no.7
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    • pp.1326-1334
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    • 2008
  • The prime-boost vaccination with DNA vaccine and recombinant viral vector has emerged as an effective prophylactic strategy to control infectious diseases. Here, we compared the protective immunities induced by multiple alternating immunizations with DNA vaccine (pCIgB) and replication-incompetent adenovirus (Ad-gB) expressing glycoprotein gB of pseudorabies virus (PrV). The platform of pCIgB-prime and Ad-gB-boost induced the most effective immune responses and provided protection against virulent PrV infection. However, priming with pCIgB prior to vaccinating animals by the DNA vaccine-prime and Ad-boost protocol provided neither effective immune responses nor protection against PrV. Similarly, boosting with Ad-gB following immunization with DNA vaccine-prime and Ad-boost showed no significant responses. Moreover, whereas the administration of Ad-gB for primary immunization induced Th2-type-biased immunity, priming with pCIgB induced Th1-type-biased immunity, as judged by the production of PrV-specific IgG isotypes and cytokine IFN-$\gamma$. These results indicate that the order and injection frequency of vaccine vehicles used for heterologous prime-boost vaccination affect the magnitude and nature of the immunity. Therefore, our demonstration implies that the prime-boost protocol should be carefully considered and selected to induce the desired immune responses.

Replication Inhibition and Its Recovery/Process in Chinese Hamster Ovary Cells Treated with Methyl Methanesulfonate (Chinese Hamster Ovary세포에 있어 methyl methanesulfonate에 의한 DNA 복제억제와 이의 회복경로)

  • 이천복;이형호;박상대;이치건
    • Environmental Mutagens and Carcinogens
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    • v.9 no.1
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    • pp.33-46
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    • 1989
  • 본 연구는 알킬화제를 처리한 CHO-K1 세포에서 DNA 복제억제와 그 회복과정의 분자론적 기작을 규명할 목적으로 방사선 이중 표지에 의한 DNA 합성율의 측정, 알칼리 자당 농도구배 초원심분리법에 의한 DNA 분자량과 후복제 회복율을 측정하여 다음과 같은 결과를 얻었다. (1) 1mM methyl methanesulfonate (MMS)와 1nM N-methyl-N'-nitro-N-nitrosoguanidine (MNNG) 이하의 낮은 농도의 처리군에서는 DNA 합성율이 급격히 감소하였으나, 2 mM MMS, 2mM MNNG이상의 농도에서는 그 감소양상이 둔화되었다, (2) DNA 합성율은 알킬화제의 처리 직후 감소하였다가 시간경과에 따라 회복되어 처리후 4시간 째에는 대조군 수준 또는 그 이상으로 회복되었다.

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On-off Regulation of 3' Exonuclease Excision to DNA Polymerization by Exo+ Polymerase

  • Zhang, Jia;Li, Kai
    • BMB Reports
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    • v.36 no.6
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    • pp.525-528
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    • 2003
  • The role of 3' exonuclease excision in DNA polymerization was evaluated in primer extensions using 3' allele-specific primers that had exonuclease-digestible and exonuclease-resistant 3' termini. With exonuclease-digestible unmodified 3' mismatched primers, the exo+ polymerase yielded template-dependent products. Using exonuclease-resistant 3' mismatched primers, no primer-extended product resulted from exo+ polymerase. As a control, polymerase without proofreading activity yielded primer-dependent products from 3' mismatched primers. These data indicated that a successful removal of the mismatch is required for DNA polymerization from the 3' mismatched primers by exo+ polymerase. In addition to the well-known proofreading from this mismatch removal, the premature termination in DNA polymerization, due to the failure of the efficient removal of the mismatched nucleotides, worked as an off-switch in maintaining the high fidelity in DNA replication from exo+ polymerase.

Characterizations of Disease Symptoms and Virus Replication Shown in the Interactions Between Arabidopsis (Arabidopsis ecotype에서 3종의 BCTV 분리주의 병증 및 복제 특성)

  • 박을용;박종범;이석찬
    • Korean Journal Plant Pathology
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    • v.14 no.5
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    • pp.507-512
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    • 1998
  • Molecular analysis has been done for characterization of the interactions between three beet curly top virus (BCTV) strains and two Arabidopsis ecotypes in terms of virus inducible disease symptoms and infectivities. The total DNA was isolated from three tissues (shoot tips, infection origins and roots) of virus infected plants and this DNA was analyzed by quantitatively and qualitatively to elucidate virus movement and symptom development. CTV-Worland infected Col-O and Sei-O showed only symptom shown in hypersusceptible ecotype Sei-O by BCTV-worland was shoot tip stunting. Kinetics of virus DNA accumulation of three different viruses indicated that roots contained more virus DNA than shoot tips or infection origins, and that disease symptom severity was strongly correlated with virus DNA accumulation. These results suggest that the mild and Worland-specific symptoms shown in Sei-O by BCTV-worland are caused by the interactions of host factors provided by hypersusceptible ecotype and viral factors of mild strain.

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Inhibition Effect of Human Cytomegalovirus Replication by Peptide nucleic acids (PNA)

  • Park, Young-Doo;Eum, Jin-Seong;Paik, Soon-Young;Hong, Seong-Karp
    • Proceedings of the Korean Institute of Information and Commucation Sciences Conference
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    • 2008.05a
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    • pp.971-975
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    • 2008
  • Human cytomegalovirus (HCMV) provokes fatal infections in AIDS patients that have deficient immune functions and patients that have cellular immune responses repressed after bone marrow transplantation. A new candidate for therapeutic against HCMV is needed because conventional treatments as ganciclovir, acyclovir, cidofovir, and foscarnet cytosine used currently are improper due to their side effects and advent of resistant HCMV. In this study, peptide nucleic acids (PNAs) against UL54 (DNA polymerase) and UL97 (phosphotransferase) that were essential in replication of HCMV were applied in inhibition of replication of HCMV. From the results of this study, 4 PNAs $_{PNA}UL97-1$, $_{PNA}UL97-2$, $_{PNA}UL54-3$, and $_{PNA}UL54-4$ showed 3.7, 3.1, 1.7, and 1.6 folds of inhibition effect against replication of HCMV in the human fibroblast cells. These PNA suggest a novel possibility as therapeutic against HCMV.

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Phylogenetic Analysis of Pleurotus Species Based on the Nuclear SSU rRNA Sequences (Phylogenetic Analysis of Pleurotus Species Based on the Nuclear SSU rRNA Sequences)

  • Jeong, Jae Hun;Kim, Eun Gyeong;No, Jeong Hye
    • Journal of Microbiology
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    • v.34 no.1
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    • pp.37-37
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    • 1996
  • The internal regions of nuclear small subunit rRNA from 6 plaeurotus species and 5 Pleurotus ostreatus strains were amplified by PCR and sequenced. The DNA sequences of 8 Pleurotus strains (P. ostreatus NFFA2, NFFA4501, NFFA4001, KFFA4001, KFCC11635, P florida, P. florida, P. sajor-cuju, P. pulmonarius, and P. spodoleucus) were idential, but P. cornucopiae differed from them in two bases out of 605 bases. However, p[hylogenetic analysis of the sequences by DNA-distance matrix and UPGMA methods showed that P. ostreatus NFFA2m1 and NFFA2m2, known as mutants of P. ostreatus NFFA2, belonged to anther group of Basidiomycotina, which is close to the genus Auricularia. The difference of the SSU rDNA sequences of P. cornucopiae from other Pleurotus species tested corresponds to the difference of mitochondrial plasmid type present in Pleurotus species as observed by Kim et al. (1993, Korean J. Microbiol. 31, 141-147).ishement of silencing at the HMR/hsp82 locus can occur in G1-arrested cells. Cell cycle arrest at G1 phase was achieved by treatment of early log a cell cultures with .alpha.-factor mating pheromone, which induces G1 arrest. The result suggests that passage through S phase (and therefore DNA replication) is nor required for re-establishing silencer-mediated repression at the HMNRa/HSP82 locus. Finally, to test whether de nono protein synthesis is required for re-establishment of silencer-mediated repression, cells were pretreated with cycloheximide (500 /.mu.g/ml) 120 min. It was apparent that inhibiting protein synthesis delays, but does not prevent, re-establishment of silencer-mediated repression. Altogether, these results indicate that re-establishment of silencer-mediated repression is not dependent on the DNA replication and has no requirement for protein synthesis.