• Title/Summary/Keyword: cdc 돌연변이

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Isolation and Characterization of the Mutants in the Genes Involved in Mating Pheromone Signalling (효모의 mating pheromone 신호전달과정에 관여하는 유전자의 돌연변이 분리 및 분석)

  • Kim, Ji-Hye;Kim, Hwan-Gyu;Jahng, Kwang-Yeop
    • The Korean Journal of Mycology
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    • v.19 no.4
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    • pp.266-275
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    • 1991
  • The gene CDC70 encoding the${\alpha}-subunit$ of G protein has been known to be a component involved in mating pheromone signalling in the yeast, Saccharomyces cerevisiae. To isolate mutations of the genes involved in the signal transduction, Saccharomyces cerevisiae the strain bearing the cdc70-5 mutation was mutagenized to be forced to recover the ability of colony-formation at restrictive temperature, which means the new mutation can suppress the temperature sensitivity of the cdc70-5 phenotypes. Among these suppressors, $sir^-$ and $mat{\alpha}2^{-}$ mutations are excluded because of no relationship to signal transducer. And the selected suppressors were analyzed for the linkage relationships by the tetrad analysis. Out of fifteen suppressors isolated, twelve were classified into four linkage groups, designated as sga1, sga2, sga3, sga4 by the tetrad analysis. The other three genes were determined for the linkage.

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Molecular Cloning of the Gene in Schizosaccharomyces pombe Related to the CDC3 Gene in Saccharomyces cerevisiae (Saccharomyces cerevisiae의 CDC3 유전자와 유사한 Schizosaccharomyces pombe 유전자의 클로닝)

  • 김형배
    • Korean Journal of Microbiology
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    • v.31 no.3
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    • pp.197-202
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    • 1993
  • The budding yeast S. cerevisiae contains 10-nm filament ring that lies just inside the plasma memhrane in the region of the mother-bud neck. It is possihle that CDC3. CDCIO, CDCII. CDCI2 genes encode the filaments. Recently it has been shown that the CDC3 and CDCI2 gene products arc localized to [he vicinity of the neck lilaments by immunolluorescence. However. the role of the lilament ring is not clear. In order to find out the role of filament ring. I have tried to clone the similar gene in S. pomhe to the CDC3 in S. cerevisiae. Genomic library was constructed by use of $\lambda$gtll expression vector and screened with CDC3 antibodies. From sequencing data, there were more than two introns in the newly cloned gene. There was 62% homology between the part of the predicted amino acid sequence of cloned gene and CDC3 amino acid sequence.

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Effect of benzo(a)pyrene and mitomycine C on HeLa cell division cycle

  • Yu, Il-Je;Lim, Cheol-Hong;Kim, Hyo-Jung;Chung, Kyu-Hyuk;Song, Kyung-Seuk;Han, Jeong-Hee;Chung, Yong-Hyun
    • Environmental Mutagens and Carcinogens
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    • v.21 no.2
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    • pp.82-88
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    • 2001
  • Recently, there has been significant progress in understanding the control process of the cell division cycle. To investigate the influence of toxic substances on the cell cycle, the effect of benzo(a)pyrene (BAP) and mitomycine C (MMC) on synchronized HeLa cells was analyzed during the cell cycle. To synchronize the HeLa cells, 10$^{6}$ cells were grown for 1 day and then treated with 1 mM hydroxyurea for 14 h. The arrested cells were then allowed to proceed through their cell cycle by removing the hydroxyurea and resupplying a fresh medium. The arrested cells in the G1/S transition then proceeded to the S phase after 4 h, the G2/M phase after 8h, and the G1 phase after 12 h, subsequent to the resupply of a fresh medium. In the untreated HeLa cells, the p34$^{cdc2}$ kinase activity, measured using a p34$^{cdc2}$ specific peptide, peaked after 8h (G2/M) and then declined after 12 h (G1). However, treatment with 30 $\mu$M BAP delayed the peak of the p34$^{cdc2}$ kinase activity. The amount of p34$^{cdc2}$ remained unchanged in the untreated, BAP-, and MMC-treated cells throughout the cell cycle. The cyclin B level peaked after 8 h in the untreated cells, yet peaked after 10-12 h in the BAP-treated cells. There was no significant change in the cyclin B level in the MMC-treated cells.

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The Role of Heat Shock Protein 25 in Radiation Resistance

  • Lee Yoon-Jin;Lee Su-Jae;Bae Sangwoo;Lee Yun-Sil
    • Environmental Mutagens and Carcinogens
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    • v.25 no.2
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    • pp.51-59
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    • 2005
  • Overexpression of HSP25 delayed cell growth, increased the level of $p21^{waf}$, reduced the levels of cyclin D1, cylcin A and cdc2, and induced radioresistance in L929 cells. We demonstrated that extracellular regulated kinase (ERK) and MAP kinase/ERK kinase (MEK) expressions as well as their activation (phospho-forms) were inhibited by hsp25 overexpression. To confirm the relationship between ERK1/2 and hsp25-mediated radioresistance, ERK1 or ERK2 cDNA was transiently transfected into the hsp25 overexpressed cells and their radioresistance was examined. HSP25-mediated radioresistance was abolished by overexpression of ERK2, but not by overexpression of ERK1. Alteration of cell cycle distribution and cell cycle related protein expressions (cyclin D, cyclin A and cdc2) by hsp25 overexpression were also recovered by ERK2 cDNA transfection. Increase in Bc1-2 protein by hsp25 gene transfection was also reduced by subsequent ERK2 cDNA-transfection. In addition, HSP25 overexpression reduced reactive oxygen species (ROS) and increased expression of manganese superoxide dismutase (MnSOD) gene. Increased activation of NF-kB (IkB degradation) was also found in hsp25-overexpressed cells. Moreover, transfection of hsp25 antisense gene abrogated all the HSP25-mediated phenomena. To further elucidate the exact relationship between MnSOD induction and NF-kB activation, dominant negative $I-kB\alpha(I-kB\alpha-DN)$ construction was transfected to HSP25 overexpressed cells. $I-kB\alpha-DN$ inhibited HSP25 mediated MnSOD gene expression. In addition, HSP25 mediated radioresistance was blocked by $I-kB\alpha-DN$ transfection. Blockage of MnSOD with antisense oligonucleotides in HSP25 overexpressed cells, prevented apoptosis and returned the ERK1/2 activation to the control level. From the above results, we suggest for the first time that reduced oxidative damage by HSP25 was due to MnSOD-mediated down regulation of ERK1/2.

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Identification of Genes Connected with the Sensitivity to 5-FU and Cisplatin in Squamous Cell Carcinoma Cell Lines (편평세포암 세포주에서 5-FU와 Cisplatin에의 감수성과 관련된 유전자의 동정)

  • Choi, Na-Young;Kim, Ok-Joon;Lee, Geum-Sug;Kim, Byung-Gook;Kim, Jae-Hyeong;Jang, Youn-Young;Lim, Won-Bong;Chong, Min-A;Choi, Hong-Ran
    • Journal of Oral Medicine and Pain
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    • v.30 no.3
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    • pp.287-300
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    • 2005
  • Squamous cell carcinoma (SCC) in head and neck show a variability in the response to chemotherapy, even when it present with similar histological tumor type, grade, and clinical stage. The purpose of present study it to identify predictive bio-marker for the sensitivity or resistance to conventional chemotherapeutic agents, 5-fluorouracil (5-FU) and Cisplatin Oral cancer cell lines were used in present study. MTT assay was performed to evaluate the sensitivity and/or resistance to 5-FU and Cisplatin. And RT-PCR was carried out for evaluation of the mRNA expressions of various genes associated with mutation, inflammation (COX pathway), cell cycle, senescence and extracellular matrix (ECM). The molecules which are correlated with the sensitivity to 5-FU are XPA, XPC, OGG, APEX, COX-2, PPAR, Cyclin E, Cyclin B1, CDC2, hTERT, hTR, TIMP-3, TIMP-4 and HSP47. And the molecules are correlated with the sensitivity to Cisplatin are COX-1, iNOS, eNOS, PCNA, collagen 1 and MMP-9. Taken together, when choosing the appropriate chemotherpeutic agents for patients, considering the molecules which are correlated or reversely correlated is helpful to choose the resonable agents for cancer patients.