• Title/Summary/Keyword: Inducible repair

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Regulation of Wound Healing and Fibrosis by Hypoxia and Hypoxia-Inducible Factor-1

  • Ruthenborg, Robin J.;Ban, Jae-Jun;Wazir, Anum;Takeda, Norihiko;Kim, Jung-Whan
    • Molecules and Cells
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    • v.37 no.9
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    • pp.637-643
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    • 2014
  • Wound healing is a complex multi-step process that requires spatial and temporal orchestration of cellular and non-cellular components. Hypoxia is one of the prominent microenvironmental factors in tissue injury and wound healing. Hypoxic responses, mainly mediated by a master transcription factor of oxygen homeostasis, hypoxia-inducible factor-1 (HIF-1), have been shown to be critically involved in virtually all processes of wound healing and remodeling. Yet, mechanisms underlying hypoxic regulation of wound healing are still poorly understood. Better understanding of how the wound healing process is regulated by the hypoxic microenvironment and HIF-1 signaling pathway will provide insight into the development of a novel therapeutic strategy for impaired wound healing conditions such as diabetic wound and fibrosis. In this review, we will discuss recent studies illuminating the roles of HIF-1 in physiologic and pathologic wound repair and further, the therapeutic potentials of HIF-1 stabilization or inhibition.

Isolation of HRD3 gene, a homologous RAD3 gene from fission yeast Schizosaccharomyces pombe

  • Choi, In-Soon;Jin, Yong-Hwan;Park, Sang-Dai
    • Environmental Mutagens and Carcinogens
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    • v.16 no.2
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    • pp.77-82
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    • 1996
  • The RAD3 gene of Saccharomyces cerevisiae is required for excision repair and is essential for cell viability. RAD3 encoded protein possesses a single stranded DNA-dependent ATPase and DNA-RNA helicase activies. To examine the extent of conservation of structure and function of RAD3 during eukaryotic evolution, we have cloned the RAD3 homolog, HRD3, from the distantly related yeast Schizosaccharomyces pombe. Here, we report the partial cloning and characterization of HRD3 gene (Homologous of RAD3 gene) which was isolated by PCR amplification using conserved domain of Saccharomyces cerevisiae RAD3 gene. Chromosomal DNA isolated from S. pombe had similar restriction patterns to those from S. cerevisiae, as determined by Southern blot analysis. The 2. 8 kb transcript of mRNA was identified by Northern hybridization. The level of transcript did not increase upon UV-irradiation, suggesting that the HRD3 gene in S. pombe is not UV-inducible.

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Characterization of UV-Inducible Gene (UVI-180) in Schizosaccharomyces pombe (분열형 효모 Schizosaccharomyces pombe에서 자외선 유도유전자 UVI-180의 특성 연구)

  • Park, In-Soon
    • Environmental Analysis Health and Toxicology
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    • v.18 no.3
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    • pp.225-230
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    • 2003
  • 본 연구는 DNA 상해유도기작을 규명하기 위하여 하등 진핵생물인 분열형 효모 Schizosaccharomyces Pombe로부터 subtraction hybridization방법을 이용하여 자외선 유도 유전자인 UVI-180을 분리하고 그 유전자 구조와 발현양상을 조사하였다. UVI-180유전자의 발현양상을 Northern hybridization 방법으로 살펴본 결과 자외선(ultraviolet-light)조사 1시간 후에 최대의 발현 증가를 나타내었다. 반면 알킬화제인 MMS(methyl methanesulfonate)처리에 의해서는 전혀 발현이 증가되지 않았다. 이 결과 UVI-180유전자는 DNA상해에 따라 각기 다른 발현양상을 나타냄을 알 수 있었다. 유전자의 기능을 알기 위하여 null-mutant세포 주를 제조하여 그 특성을 살펴본 결과 이 유전자는 세포의 성장에 필수적인 유전자임을 알 수 있었다.

A gene responsible for ozone sensitivity (ozrB) in chromosome of escherichia coli B. MQ 1844 (E. coli B. MQ1844 균주의 오존감수성 유전자의 특성)

  • ;Hamelin C.
    • Korean Journal of Microbiology
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    • v.25 no.2
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    • pp.103-109
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    • 1987
  • An ozone-sensitive mutant of Escherichia coli strain B, MQ 1844 is described. Its properties, including high sensitivity to ozone and radiation, inducible filamentation, extensive DNA degradation and impaired DNA synthesis following ozonation, are attributable to a mutation in ozrB, a gene which is cotransducible with malB. Based on differences in phenotypic expression as well as on the particular location of this gene on the bacterial chromosome, ozrB appears as distinct from the other ozone-or radiation-sensitivity genes previously described.

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Characterization of Excision Repair Genes Related to Damaged DNA Repair from Eukaryotic Cells

  • Choi, In-Soon;Jin, Yong-Hwan;Park, Sang-Dai
    • Environmental Mutagens and Carcinogens
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    • v.17 no.1
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    • pp.1-6
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    • 1997
  • The RAD4 gene of Saccharomyces cerevisiae is essential for the incision step of UV-induced excision repair. A yeast RAD4 gene has been previously isolated by functional complementation. In order to identify the RAD4 homologous gene from fungus Coprinus cinereus, we have constructed cosmid libraries from electrophoretically separated chromosomes of the C. cinereus. The 13 C. cinereus chromosomes were resolved by pulse-field gel electrophoresis, hybridized with S. cerevisiae RAD4 DNA, and then isolated homologous C. cinereus chromosome. The insert DNA of the RAD4 homolog was contained 3.2 kb. Here, we report the partial cloning and characterization of fungus C. cinereus homolog of yeast RAD4 gene. Southern blot analysis confirmed that C. cinereus contains the sequence homologous DNA to RAD4 gene and this gene exists as a single copy in C. cinereus genome. When total RNA isolated from C. cinereus cells was hybridized with the 1.2 kb PvuII DNA fragment of the S. cerevisiae RAD4 gene, a 2.5 kb of transcript was detected. The level of the transcript did not increase upon UV-irradiation, suggesting that the RAD4 homologous gene in C. cinereus is not UV-inducible.

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Effects of Anti-Neoplastic Antibiotics on DNA Replication and Repair (DNA복제 및 회복에 미치는 수종항암 항생제의 영향에 관한 연구)

  • Park, Sang-Dai;Rie, Myung-Chull;Lee, Chun-Bok
    • The Korean Journal of Zoology
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    • v.26 no.1
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    • pp.19-28
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    • 1983
  • Alkaline elution profiles showed that the frequency of DNA single strand breaks associated with DNA-protein crosslinks in cells treated with both an inducing dose of MMC $(MMC_1)$ and a challenge dose of MMC $(MMC_2)$ was slightly less than that in cells treated with MMC alone. The amount of unscheduled DNA synthesisi in cells treated with both $MMC_1$ and $MMC_2$ was greater than that in cells treated with MMC alone. This enhancement of exicision repair detected by UDS autoradiography and alkaline elution, was not observed, when cells were incubated with cyclohexmide between the two treatments of $MMC_1$ and $MMC_2$. These results suggest that MMC-damaged DNA from Chinses hamster cells is repaired by excision repair mechanisms that require de novo protein synthesis for enhancement, and that an inducible repair mechanism may exist in CHO cells.

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Unbalanced Restriction Impairs SOS-induced DNA Repair Effects

  • Katna, Anna;Boratynski, Robert;Furmanek-Blaszk, Beata;Zolcinska, Natalia;Sektas, Marian
    • Journal of Microbiology and Biotechnology
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    • v.20 no.1
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    • pp.30-38
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    • 2010
  • The contribution of a type II restriction-modification system (R-M system) to genome integrity and cell viability was investigated. We established experimental conditions that enabled the achievement of hemimethylated and unmethylated states for the specific bases of the recognition sequences of the host's DNA. To achieve this, we constructed the MboII R-M system containing only one (i.e., M2.MboII) out of two functional MboII methyltransferases found in Moraxella bovis. Using the incomplete R-M system, we were able to perturb the balance between methylation and restriction in an inducible manner. We demonstrate that upon the SOS-induced DNA repair in mitomycin C treated cells, restriction significantly reduces cell viability. Similar results for the well-studied wild-type EcoRI R-M system, expressed constitutively in Escherichia coli, were obtained. Our data provide further insights into the benefits and disadvantages of maintaining of a type II R-M system, highlighting its impact on host cell fitness.

Characterization of RAD4 Homologous Gene from Coprinus cinereus (균류 Coprinus cinereus에서 DNA 회복에 관여하는 RAD4 유사유전자의 분리와 특성)

  • Choi, In-Soon
    • Journal of Life Science
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    • v.13 no.4
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    • pp.522-528
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    • 2003
  • The RAD4 gene of Saccharomyces cerevisiae is essential for the incision step of UV-induced excision repair. A yeast RAD4 gene has been previously isolated by functional complementation. In order to identify the RAD4 homologous gene from fungus Coprinus cinereus, we have constructed cosmid libraries from electrophoretically separated chromosomes of the C. cinereus. The 13 C. cinereus chromosomes were resolved by pulse-field gel electrophoresis, hybridized with S. cerevisiae RAD4 DNA, and then isolated homologous C. cinereus chromosome. The insert DNA of the RAD4 homolog was contained 3.2 kb. Here, we report the characterization of fungus C. cinereus homolog of yeast RAD4 gene. Southern blot analysis confirmed that C. cinereus contains the RAD4 homolog gene and this gene exists as a single copy in C. cinereus genome. When total RNA isolated from C. cinereus cells was hybridized with the 1.2 kb PvuII DNA fragment of the S. cerevisiae RAD4 gene, a 2.5 kb of transcript was detected. In order to investigation whether the increase of transcripts by DNA damaging agent, transcripts levels were examined after treating the cells. The level of transcript did not increase by untraviolet light (UV). This result indicated that the RAD4 homologous gene is not UV inducible gene. Gene deletion experiments indicate that the RAD4 homologous gene is essential for cell viability.

Characterization of RAD3 Homologous Gene from Coprinus cinereus (균류 Coprinus cinereus에서 DNA 회복에 관여하는 RAD3 유사유전자의 분리와 특성)

  • Choi In Soon
    • Journal of Life Science
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    • v.14 no.6 s.67
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    • pp.1023-1027
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    • 2004
  • The RAD3 gene of Saccharomyces cerevisiae is essential for the incision step of UV-induced excision repair. An yeast RAD3 gene has been previously isolated by functional complementation. In order to identify the RAD3 homologous gene from fungus Coprinus cinereus, we have constructed cosmid libraries from electrophoretically separated chromosomes of the C. cinereus. The 13 C. cinereus chromosomes were resolved by pulse-field gel electrophoresis, hybridized with S. cerevisiae RAD3 DNA, and then isolated RAD3 homologous DNA from C. cinereus chromosome. The RAD3 homolog DNA was contained in 3.2 kb DNA fragment. Here, we report the results of characterization of a fungus C. cinereus homolog to the yeast RAD3 gene. Southern blot analysis confirmed that the C. cinereus chromosome contains the RAD3 homolog gene and this gene exists as a single copy in C. cinereus genome. When total RNA isolated from the C. cinereus cells were hybridized with the 3.4 kb PvuII DNA fragment of the S. cerevisiae RAD3 gene, transcripts size of 2.8 kb were detected. In order to investigate whether the increase of the amount of transcripts by DNA damaging agent, transcript levels were examined after treating agents to the cells. The level of transcripts were not increased by untraviolet light (UV). This result indicated that the RAD3 homologous gene is not UV inducible gene. Gene deletion experiments indicate that the HRD3 gene is essential for viability of the cells and DNA repair function. These observations suggest an evolutionary conservation of other protein components with which HRD3 interacts in mediating its DNA repair and viability functions.

A Study for Regulation of Ethanol-inducible $P_{450}$(CYP2E1) on $CCI_4$-induced Hepatic Damage

  • Park, Sun-Mi;Park, Eun-Jeon;Ko, Geon-Il;Kim, Jae-Baek;Sohn, Dong-Hwan
    • Archives of Pharmacal Research
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    • v.18 no.3
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    • pp.179-182
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    • 1995
  • Previous study showed that $CCl_4$ administration evoked a rapid decrease in cytochrome $P_{450}$ 2E1 protein soon after the exposure due to posttranslational inhibition(Biochem. Biophys. Res. Commun. 179:449-454, 1991). In this report, aniline hydroxylase and the amounts of immunoreactive $P_{450}$ 2E1 were rapidly decreased during day 1 to 2 and recovered during day 3 to 4 after a single dose of $CCl_4$. The activity of pentoxyresorufin-O-dealkylase was also suppressed at day 1 and began to repair from day 2. However, the decrease in immunoreactive $P_{450}$ 2C content was not observed. The decreases in $P_{450}$ 2E1 enzyme activity and immunoreactive protein by acute $CCl_4$ treatment were accompanied by a decline in $P_{450}$ 2E1 mRNA level. The data thus suggested a pretranslational reduction of $P_{450}$ 2E1 during day 1 to 2 after acute $CCl_4$ treatment.

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