• Proceedings of the Korean Society of Life Science Conference
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• 2005.09a
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• pp.37-37
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• 2005

• Journal of Photoscience
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• v.7 no.2
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• pp.53-58
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• 2000

We performed the cloning of a chalcone synthase (CHS) gene, the key enzyme in the anthocyanin biosynthesis, from the cDNA library constructed with grape suspension cells irradiated UV-B. The PCR fragment was used to cloning the CHS gene. One CHS cDNA clone containing an open reading frame and a partial stilbene synthase (STS)cDNA, the stilbene-type phytoalexin, were isolated. The CHS cDNA clone (VCHS) showed 87% sequence homology with VvCHS (V.vinifea) and 72.3% identity with VSTSY(V.vinifea). its amino acid sequences were longer than any other CHS genes as 454 residues. Two genes were weakly expressed in white light irradiated cells, but highly induced in UV-B irradiated condition during 32 hours. Interestingly, the STS was quickly and abundantly expressed from 2 hours when supplemented with jasmonic acid (JA) and the maximum expression was observed at 4 hours and then gradually decreased. But, the additional UV-B or white light quickly degraded the STS expression than only JA treated grape suspension cells. The CHS also was rapidly induced with JA and the synergistical effect was observed at the addigional light treatment of UV-B or white light. These results are indicated that CHS and STS have different response mechanisms against the environmental stresses.

• 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.

• Proceedings of the Zoological Society Korea Conference
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• 1996.10a
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• pp.225.2-225
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• 1996

No Abstract, See Full Text

• Animal cells and systems
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• v.7 no.2
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• pp.159-164
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• 2003

The RAD3 gene of Saccharomyces cerevisiae is required for excision repair and is essential for cell viability. The RAD3 encoded protein possesses a single stranded DNA-dependent ATPase and DNA and DNA-RNA helicase activities. To examine the extent of conservation of structure and function of a S. pombe RAD3 during eukaryotic evolution, the RAD3 homolog gene was isolated by screening of genomic DNA library. The isolated gene was designated as HRD3 (homolog of RAD3 gene). Southern blot analysis confirmed that S. pombe chromosome contains the same DNA as HRD3 gene and this gene exists as a single copy in S. pombe. The transcript of 2.8 kb was detected by Northern blot analysis, The level of transcripts increased by ultraviolet (UV) irradiation, indicating that HRD3 is one of the UV-inducible genes in S. pombe. Furthermore, the predicted partial sequence of HRD3 protein has 60％ identity to S. cerevisiae RAD3 gene. This homology was particularly striking in the regions identified as being conserved in a group of DNA helicases. Gene deletion experiments indicate that the HRD3 gene is essential for viability and DNA repair function. These observations suggest evolutionary conservation of other protein components with which HRD3 might interact in mediating its DNA repair and viability functions.

• Journal of Plant Biotechnology
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• v.29 no.2
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• pp.79-84
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• 2002

Suppression subtractive hybridization (SSH) was used to isolate wound-induced cDNAs from wounded soybean. One of wound-induced cDNA, gmwi33 showed high homology with genes encoding $\beta$-amyrin synthase. The full length cDNA of gmwi33, designated GmAMS1, is 2416 bp long and contains an open reading frame consisted of 739 amino acids. GmAMS1 protein showed 89% identity with licorice GgbAS1 and 86% identity with pea OSCPSY. In 5 day-old, dark-grown seedlings, the expression of GmAMS1 was most strongly induced by light and weakly induced by methyl jasmonate and by low temperature. However, GmAMS1 was not induced by elicitor or UV-B treatment. Such expression pattern might be closely related with the oxygen-radical scavenging activity of soyasaponin.

• The Korean Journal of Zoology
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• v.37 no.2
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• pp.232-239
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• 1994

RecA protein plans a central role in homologous recombination and DNA repair in Escherichia cofi (E. colD. The function 8nd structure of this protein are universal in prokarvotes and also conserved in eukaryotes such as yeast. The RecA-like protein with 74 lInDa in size has already been identified and purified from a fission yeast Schizosaccharomyces pombe (5. pommel (Lee, 19911. From this study it was revealed that the RecA-like protein of 5. pombe was highly inducible to various DNA damaging agents and inhibitors of nucleotide pool svnthesizins enzymes. The cellular level of the 5. pombe RecA-like protein wi,u markedly increased, upto 5- to 10-fold, by treatment with various DNA-damains agents including ultraviolet (UV) light, methyl methanesulfonate WS),4-nitroquinoline-1-oxide (4-NQO), and mitomycin-C (MMC), similar to E. cofi RecA protein. Interestingly, the protein level was also increased by inhibitors of nucleotide pool forming enzlwnes such as methotrexate (MTX) and hvdroxvurea (HU). The most effective doses for the inducibility of 4-NQO, MMS, W, MMC, MTX, and HU were 0.2 Ug/ml, 30 mM, 200 J/ma, 0.4 $\mus/ml,$ 1 Ug/ml, and 100 mM, respectively. The range of effective duration time for the inducibilitv of RecA-like protein was from 270 to 450 mins. These results suggest that the 5. pombe RecA-like protein also platys an imortant role in cellular responses to DNA damage as in E. coli system.

• Animal cells and systems
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• v.3 no.2
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• pp.229-236
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• 1999

The recA gene plays a central role in genetic recombination and SOS DNA repair in Escherichia coli (E. coli). We have previously identified a 42 kDa RecA-like protein inducible by a variety of DNA damages from a fluorescent Pseudomonas strain sp. and characterized its inducible kinetics. In the present study, we cloned and characterized the gene encoding the RecA-like protein by immunological screening of Pseudomonas genomic expression library using polyclonal E. coli anti-RecA antibodies as a probe. From 10$^{5}$ plaques screened, five putative clones were finally isolated. Southern blot analysis indicated that four clones had the same DNA inserts and the recA-like gene was located within the 3.2 kb EcoRI fragment of Pseudomonas chromosomal DNA. In addition, the cloned recA-like gene was transcribed into an RNA transcript approximately 1.1 kb in size, as judged by Northern blot analysis. The cellular level of RNA transcript of the cloned recA-like gene was increased to an average of 5.15- fold upon treatment with DNA damaging agents such as ultraviolet (UV)- light, nalidixic acid (NA), methyl methanesulfonate (MMS), and mitomycin-C (MMC). These results suggest that the cloned gene is inducible by DNA damage similarly to the recA gene in E. coli. However, the cloned gene did not restore the DNA damage sensitivity of the E. coli recA-mutant.

• Environmental Mutagens and Carcinogens
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• v.22 no.3
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• pp.137-141
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• 2002

The SNF2/SW12 family comprises proteins from a variety of species with in vivo functions, such as transcriptional regulation, maintenance of chromosome stability during mitosis, and various types of DNA repair. This study was shown the characterization of hrp2+ gene which was isolated by PCR amplification using the conserved domain of SNF2 motifs. Sequence analysis of hrp2+ gene showed striking evolutionary conservation among the SNF2 family of proteins. The transcript of hrp2+ gene was found to be a 4.7 kb as identified by Northern hybridization. In addition, to determine the transcription initiation site of hrp2+ gene, primer extension analysis was performed. This result showed the band of 64 bp. The transcriptional start point was mapped to a position of 47 base pair from the first ATG codon of translational initiation codon. In order to investigate the inducibility of hrp2+ gene, transcript levels were examined after treating the cells to various DNA damaging agents. The transcripts of hrp2+ were induced by UV-irradiation. But the transcripts were not induced by treatment of 0.25% Methylmethane sulfonate (MMS). These results implied that the effects of damaging agents are complex and different regulatory pathways exist for the induction of this gene.

• Journal of Life Science
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• v.15 no.2 s.69
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• pp.192-196
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• 2005

The SNF2/SW12 family comprises proteins from a variety of species with in vivo functions, such as transcriptional regulation, maintenance of chromosome stability during mitosis, and various types of DNA repair. This study was shown the characterization of hrp2+ gene which was isolated by PCR amplification using the conserved domain of SNF2 motifs. Sequence analysis of hrp2+ gene showed striking evolutionary conservation among the SNF2 family of proteins. The transcript of hrp2+ gene was found to be a 4.7 kb as identified by Northern hybridization. To investigate the inducibility of hrp2+ gene, transcript levels were examined after treating the cells to various DNA damaging agents. The transcripts of hrp2+ were induced by UV-irradiation. But the transcripts were not induced by treatment of $ 0.25\%$ Methylmethane sulfonate (MMS). These results implied that the effects of damaging agents are complex and different regulatory pathways exist for the induction of this gene. Hrp2 protein was purified near homogeneity by combination of affinity chromatography. We tested the purified Hrp2 protein for the helicase activity in an oligonucleotide release assay. However we were unable to detect any helicase activity associated with the Hrp2 protein, indicating that the helicase motifs in Hrp2 are merely indicators of a broader DNA-dependent ATPase activity.