• Title/Summary/Keyword: Natural DNA

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Phloroglucinol Attenuates Ultraviolet B-Induced 8-Oxoguanine Formation in Human HaCaT Keratinocytes through Akt and Erk-Mediated Nrf2/Ogg1 Signaling Pathways

  • Piao, Mei Jing;Kim, Ki Cheon;Kang, Kyoung Ah;Fernando, Pincha Devage Sameera Madushan;Herath, Herath Mudiyanselage Udari Lakmini;Hyun, Jin Won
    • Biomolecules & Therapeutics
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    • v.29 no.1
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    • pp.90-97
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    • 2021
  • Ultraviolet B (UVB) radiation causes DNA base modifications. One of these changes leads to the generation of 8-oxoguanine (8-oxoG) due to oxidative stress. In human skin, this modification may induce sunburn, inflammation, and aging and may ultimately result in cancer. We investigated whether phloroglucinol (1,3,5-trihydroxybenzene), by enhancing the expression and activity of 8-oxoG DNA glycosylase 1 (Ogg1), had an effect on the capacity of UVB-exposed human HaCaT keratinocytes to repair oxidative DNA damage. Here, the effects of phloroglucinol were investigated using a luciferase activity assay, reverse transcription-polymerase chain reactions, western blot analysis, and a chromatin immunoprecipitation assay. Phloroglucinol restored Ogg1 activity and decreased the formation of 8-oxoG in UVB-exposed cells. Moreover, phloroglucinol increased Ogg1 transcription and protein expression, counteracting the UVB-induced reduction in Ogg1 levels. Phloroglucinol also enhanced the nuclear translocation of nuclear factor erythroid 2-related factor 2 (Nrf2) as well as Nrf2 binding to an antioxidant response element located in the Ogg1 gene promoter. UVB exposure inhibited the phosphorylation of protein kinase B (PKB or Akt) and extracellular signal-regulated kinase (Erk), two major enzymes involved in cell protection against oxidative stress, regulating the activity of Nrf2. Akt and Erk phosphorylation was restored by phloroglucinol in the UVB-exposed keratinocytes. These results indicated that phloroglucinol attenuated UVB-induced 8-oxoG formation in keratinocytes via an Akt/Erk-dependent, Nrf2/Ogg1-mediated signaling pathway.

Opposite Effect of Spermine on the Susceptibility of Native and Denatured Calf Thymus DNA to DNase 1. (Native 및 Denatured Calf Thymus DNA의 DNase 1 에 대한 Susceptibility에 미치는 Spermine의 영향)

  • Thong-Sung Ko;Joon Huh;Chun-Bae Lee;Moo- Kyeu Park
    • Journal of the Korean Chemical Society
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    • v.27 no.6
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    • pp.429-433
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    • 1983
  • Spermine has opposite effect on the susceptibility of native and denatured calf thymus DNA toward deoxyribonuclease Ⅰ (DNase I; deoxyribonucleate 5'-oligonucleotidohydrolase; EC 3.1.4.5; from bovine pancreas). It enhances the susceptibility of the native DNA, whereas depresses that of the denatured DNA. In the absence of spermine, the reactivity of the deoxyribonuclease is apparently insensitive to the conformational difference of the native and denatured DNA.

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Analysis of Genetic Variation in Botrytis cinerea Isolates Using Random Amplified Polymorphic DNA Markers

  • Choi, In-Sil;Kim, Dae-Hyuk;Lee, Chang-Won;Kim, Jae-Won;Chung, Young-Ryun
    • Journal of Microbiology and Biotechnology
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    • v.8 no.5
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    • pp.490-496
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    • 1998
  • Random amplified polymorphic DNA (RAPD) markers were used to survey genetic variability among 34 Botrytis cinerea isolates from nine different host plants in Korea. For RAPD analysis, 115 arbitrary decamer primers were initially screened for polymorphic major DNA bands with 11 representative B. cinerea isolates. Eleven primers that initially detected polymorphisms were tested a second time with additional 23 isolates of B. cinerea as well as one isolate of Botrytis squamosa as an outgroup. The RAPD analyses revealed that all isolates except one showed different molecular phenotypes. Dendrograms obtained from dissimilarity matrices using the unweighted paired group method of arithmetic means (UPGMA) showed the 36.4% to 90.0% similarity among all B. cinerea isolates. The B. squamosa isolate showed the least similarity to all B. cinerea isolates. The cluster analyses indicated no correlation among all the characteristics examined including molecular phenotypes, host and geographic origins, year of isolation, or pathogenicity. The RAPD data suggest that a high level of genetic variation exists among Korean populations of B. cinerea and it seems to be caused by heterokaryosis among preexisting molecular phenotypes.

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OxyR Regulon Controls Lipid Peroxidation-mediated Oxidative Stress in Escherichia coli

  • Yoon, Seon-Joo;Park, Ji-Eun;Yang, Joon-Hyuck;Park, Jeen-Woo
    • BMB Reports
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    • v.35 no.3
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    • pp.297-301
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    • 2002
  • Membrane lipid peroxidation processes yield products that may react with DNA and proteins to cause oxidative modifications. The oxyR gene product regulates the expression of enzymes and proteins that are needed for cellular protection against oxidative stress. Upon exposure to tert-butylhydroperoxide (t-BOOH) and 2,2'-azobis (2-amidinopropane) hydrochloride (AAPH), which induce lipid peroxidation in membranes, the Escherichia coli oxyR overexpression mutant was much more resistant to lipid peroxidation-mediated cellular damage, when compared to the oxyR deletion mutant in regard to growth kinetics, viability, and DNA damage. The deletion of the oxyR gene in E. coli also resulted in increased susceptibility of superoxide dismutase to lipid peroxidation-mediated inactivation. The results indicate that the peroxidation of lipid is probably one of the important intermediary events in free radical-induced cellular damage. Also, the oxyR regulon plays an important protective role in lipid peroxidation-mediated cellular damage.

Cloning of Genomic DNAs of Trametes versicolor Acting as Autonomously Replicating Sequences in Saccharomyces cerevisiae

  • Sora An;Park, Kyoung-Phil;Park, Hyoung-Tae;Kim, Kyu-Joong;Kim, Kyunghoon
    • Journal of Microbiology
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    • v.40 no.3
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    • pp.245-247
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    • 2002
  • A genomic DNA library of the fungus Trametes versicolor was constructed in a yeast integration vector which contains the URA3 gene of the budding yeast Saccharomyces cerevisiae and the gene responsible for hygromycin B resistance, and fragments acting as autonomously replicating sequences (ARSes) in the budding yeast were identified from the genomic DNA library. Sixteen recombinant plasmids from the library transformed the budding yeast Saccharomyces cerevisiae to Ura+ at high frequencies. They were maintained stably under selective conditions, but were gradually lost from yeast cells at different rates under nonselective conditions, indicating that they contain eukaryotic origins of DNA replication and exist as extrachromosomal plasmids. Base sequences of four ARS DNAs among the 16 cloned fragments revealed that all or the four contain at least one 11 bp [(A/T)TTTA(T/C)(A/G)TTT(A/T)]consensus sequence of the budding yeast ARS.

Molecular Genetic Analysis of Ancient Cattle Bones Excavated from Archaeological Sites in Jeju, Korea

  • Kim, Jae-Hwan;Oh, Ju-Hyung;Song, Ji-Hoon;Jeon, Jin-Tae;Han, Sang-Hyun;Jung, Yong-Hwan;Oh, Moon-You
    • Molecules and Cells
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    • v.20 no.3
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    • pp.325-330
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    • 2005
  • Ancient cattle bones were excavated from archaeological sites in Jeju, Korea. We used molecular genetic techniques to identify the species and establish its relationship to extant cattle breeds. Ancient DNA was extracted from four sources: a humerus (Gonae site, A.D. 700-800), two fragments of radius, and a tooth (Kwakji site, A.D. 0-900). The mitochondrial DNA (mtDNA) D-loop regions were cloned, sequenced, and compared with previously reported sequences of various cattle breeds (9 Asian, 8 European, and 3 African). The results revealed that these bones were of the breed, Bos taurus, and a phylogenetic tree indicated that the four cattle bones formed a monophyletic group with Jeju native black cattle. However, the patterns of sequence variation and reports from archaeological sites suggest that a few wild cattle, with a different maternal lineage, may have existed on Jeju Island. Our results will contribute to further studies of the origin of Jeju native cattle and the possible existence of local wild cattle.

Expression of Thiol-Dependent Protector Protein from Yeast Enhances the Resistance of Escherichia coli to Menadione

  • Park, Jeen-Woo;Ahn, Soo-Mi;Kim, Eun-Ju;Lee, Soo-Min
    • BMB Reports
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    • v.29 no.6
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    • pp.513-518
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    • 1996
  • A soluble protein from Saccharomyces cerevisiae specifically provides protection against a thiolcontaining oxidation system but not against an oxidation system without thiol. This 25-kDa protein was thus named thiol-dependent protector protein (TPP). The role of TPP in the cellular defense against oxidative stress was investigated in Escherichia coli containing an expression vector with a yeast genomic DNA fragment that encodes TPP (strain YP) and a mutant in which the catalytically essential amino acid in the active site of TPP (Cys-47) has been replaced with alanine by site-directed mutagenesis (strain YPC47A). There was a distinct difference between these two strains in regard to viability, modulation of activities of superoxide dismutase and catalase, and the oxidative damage of DNA upon exposure to menadione. These results suggest that TPP may play a direct role in the cellular defense against oxidative stress by functioning as an antioxidant protein.

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Sirt1 Promotes DNA Damage Repair and Cellular Survival

  • Song, Seung-Hyun;Lee, Mi-Ok;Lee, Ji-Seon;Oh, Je-Sok;Cho, Sung-Uk;Cha, Hyuk-Jin
    • Biomolecules & Therapeutics
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    • v.19 no.3
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    • pp.282-287
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    • 2011
  • Sirt1, a nicotinamide adenine dinucleotide ($NAD^+$)-dependent histone deacetylase, is known to deacetylate a number of proteins that are involved in various cellular pathways such as the stress response, apoptosis and cell growth. Modulation of the stress response by Sirtuin 1 (Sirt1) is achieved by the deacetylation of key proteins in a cellular pathway, and leads to a delay in the onset of cancer or aging. In particular, Sirt1 is known to play an important role in maintaining genomic stability, which may be strongly associated with a protective effect during tumorigenesis and during the onset of aging. In these studies, Sirt1 was generated in stably expressing cells and during the stimulation of DNA damage to examine whether it promotes survival. Sirt1 expressing cells facilitated the repair of DNA damage induced by either ionizing radiation (IR) or bleomycin (BLM) treatment. Fastened damaged DNA repair in Sirt1 expressing cells corresponded to prompt activation of Chk2 and ${\gamma}$-H2AX foci formation and promoted survival. Inhibition of Sirt1 enzymatic activity by a chemical inhibitor, nicotinamide (NIC), delayed DNA damage repair, indicating that promoted DNA damage repair by Sirt1 functions to induce survival when DNA damage occurs.