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Choi, Tae-Yeong,Cho, Nam-Young,Oh, Younsang,Yoo, Mi-Ae,Matsukage, Akio,Ryu, Yoonseok,Han, Kyuhyung,Yoon, Jaeseung,Baek, Kwanghee 경희대학교 생명자원과학연구원 2000 硏究論文集 Vol.21 No.-
The TATA box binding protein (TBP) is a general transcription factor required for initiation by all three eukaryotic RNA polymerases. Previously, we found that the promoter region of the Drosophila melanogaster TBP gene contains three sequences similar to the DNA replication-related element (DRE) (5-TATCGATA). In the present study, we found that the DRE-like sequences are also present in the promoter of the Drosophila virilis TBP gene, suggesting a role for these sequences in TBP expression. Band mobihty shift assays revealed that oligonucleotides containing sequences similar to the DRE of D. melanogaster TBP gene promoter form specific complexes with a factor in a Kc cell nuclear extract and with recombinant DRE-binding factor (DREF). Furthermore, these complexes were either supershifted or diminished by monoclonal antibodies to DREF. Transient luciferase assays demonstrated that induction of mutations in two DRE-related sequences at positions -223 and -63 resulted in an extensile reduction of promoter activity. Thus, the DRE-DREF system appears to be involved in the expression of the D. melanogaster TBP gene. ⓒ 2000 Federation of European Biochemcal Societies. Published by Elsevier Science B.V. All rights reserved.
Ryu, Jae-Ryeon,Choi, Thae-Yeong,Kwon, Eun-Jeong,Lee, Won-Ho,Nishida, Yasuyoshi,Hayashi, Yuko,Matsukage, Akio,Yamaguchi, Masamitsu,Yoo, Mi-Ae 부산대학교 유전공학연구소 1997 분자생물학 연구보 Vol.13 No.-
The DRE/DREF system plays an important role in transcription of DNA replication genes such as those encoding the 180 and 73 kDa subunits of DNA polymeraseαas well as that encoding PCNA. In this study, we found two sequences homologous to DRE(5'-TATCGATA-3')in the 5'-flanking region(-370 to-357 with respect to the transcription initiation site) of the D-raf gene and confirmed transcription activity through gel mobility shift assay, transient CAT assays, and spatial patterns of lacZ expression in transgenic larval tissues carrying D-raf gene is another target of the Zerknu¨llt(Zen)protein with observation of D-raf repression by Zen protein in cultured cells anf its ectopic expression in the dorsal region of the homozygous zen mutant embryo. The evdience of DRE/DREF involvement in regulation of the D-raf gene obtained in this study strongly supports the idea that the DRE/DREF system is responsible for the coordinated regulation of cell proliferation-related genes in Drosophila.
Yoo, Mi-Ae,LEE, Won-Ho,HA, Hye-Yeong,RYU, Jae-Ryeon,YAMAGUCHI, Masamitsu,FUJIKAWA, Kazuo,MATSUKAGE, Akio,KONDO, Sohei,NISHIDA, Yasuyoshi 부산대학교 유전공학연구소 1994 분자생물학 연구보 Vol.10 No.-
DNA polymerase β(pol β)cDNA of rat fused to an enhamcer-promoter ergion plus a poly(A) signal sequence of actin 5C gene of Drosophila(abbreviated polβ) was transferred to the Drosophila genome. Three of four constructed transgenic strains possessing transgene polβ on different chromosomes were studied. Levels of the polβ transcript and those of the polymerization activity of polβ were markedly elevated in cultured cells trans fected with polβ-bearing vectors as well as in embryos of the transgenic strains. The popular idea that DNA polymerase β participates in DNA repair was not supported by the observation that a pair of a normal and a polβ strain, and the other pair of a mei-9 mei-41(DNA-repair deficient double mutations) strain and a polβmei-9 mei-41 strain, showed no difference in survial within each pair after treatment with ultraviolet light, methylmethane sulfonate and mitomycin C. The other idea that DNA polymerase β participates in recombination was supported by the findings that spontaneous freauency of recombination, either meiotic or mitotic, is significantly higher in a transgenic polβstrain than in a non-transgenic strain. The enhanced recomdination frequency in the polβ strain may, however, reflect an indirect effect of over-produced polβ prteins on chromosomal stability. Whatever the direct effect of rat polβis, the transgenic polβ flies will be useful for study of the physiological role of polβ and the mechanism of recombination.
E2F-dependent transcription of the rafproto-oncogene during Drosophila development
Kwon, Eun-Jeong,Oh, Eun-Jin,Kim, Young-Shin,Hirose, Fumiko,Ohno, Katsuhito,Nishida, Yasuyoshi,Matsukage, Akio,Yamaguchi, Masamitsu,Yoo, Mi-Ae 부산대학교 유전공학연구소 2001 분자생물학 연구보 Vol.17 No.-
D-raf, a Drosophila homolog of the raf proto-oncogene, has diverse functions throughout development and is transcribed in a wide range of tissues, with high levels of expression in the ovary and in association with rapid proliferation. The expression pattern resembles those of S phase genes, which are regulated by E2F transcription factors. In the 5'-flanking region of D-raf, four sequences (E2F sites 1-4) similar to the E2F recognition sequence were found, one of them (E2F site 3) being recognized efficiently by Drosophila E2F (dE2F) in vitro. Transient luciferase expression assays confirmed activation of the D-raf gene promoter by dE2F/dDP. Expression Eraf-lacZ was greatly reduced in embryos homozygous for the dE2F mutation. These results suggest that dE2F is likely to be an important regulator of D-raf transcription.
Kwon, Eun-Jeong,Park, Hyun-Sook,Kim, Young-Shin,Oh, Eun-Jin,Nishida, Yasuyoshi,Matsukage, Akio,Yoo, Mi-Ae,Yamaguchi, Masamitsu 부산대학교 유전공학연구소 2000 분자생물학 연구보 Vol.16 No.-
The Drosophila raf (D-raf) gene promoter contains a recognition consensus sequence for Drosophila STAT (D-STAT). By band mobility shift assay, we detected a factor binding to the D-STAT-recognition sequence in extracts of cultured Drosophila cells treated with vanadate peroxide. UV-cross-linking analyses suggested the size of the binding factor to be almost same as that of D-STAT. Furthermore, the binding activity was increased in cells cotransfected with HOP and D-STAT expression plasmids. These results strongly suggest that D-STAT binds to the D-STAT recognition sequence in the D-raf gene promoter. Transient luciferase expression assay using Schneider 2 cells indicated that the D-raf gene promoter is activated by D-STAT through the D-STAT-binding site. Furthermore, analyses with transgenic flies carrying Draf-lacZ gene promoter activity throughout development. We also found that the D-STAT-binding site is required for injury-induced activation of the D-raf gene promoter. Here we propose that D-STAT can participate in regulation of the mitogen-activated protein kinase cascade through D-raf gene activation.