2000 Fiscal Year Final Research Report Summary
Tissue specific regulation of DNA replication during Drosophila development.
| Project/Area Number |
11640625
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| Research Category |
Grant-in-Aid for Scientific Research (C)
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| Allocation Type | Single-year Grants |
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| Section | 一般 |
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| Research Field |
遺伝
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| Research Institution | Kyoto Institute of Technology (2000)
Aichi Cancer Center Research Institute (1999) |
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Principal Investigator |
H.INOUE Yoshihiro Kyoto Istitute of Technology, Drosophila Genetic Res.Ctr., Lecture, ショウジョウバエ遺伝資源センター, 講師 (90201938)
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| Co-Investigator(Kenkyū-buntansha) |
YAMAGUCHI Masamitsu Aichi Cancer Ctr.Res.Inst., Section Head, 増殖制御研究室, 室長 (00182460)
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| Project Period (FY) |
1999 – 2000
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| Keywords | DNA replication / Drosophila / DNA polymerase / Pelement |
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| Research Abstract |
It has been reported that several variations of cell cycle are observed during Drosophila development. Most of larval cells undergoes endoreplication cycle which comprises repeated rounds of S phase. In contrast, the imaginal disc cells proliferate with conventional cell cycle consisted of Gl, S, G2 and M phases. We have identified a mutation for the 140 kD subunit of DNA replication factor, RF-C as a dominant suppressor of rough eye phenotypes induced by ectopic expression of DREF, a common transcription factor for many genes involved in DNA
replication. The mutant dies during larval/pupal stages because of DNA replication defects in imaginal disc cells that are mitotically proliferating, while any abnormalities in endoreplication was not evident in larval cells. Taking together that factors involved in DNA replication were supplied as maternal products, the phenotypes suggests an idea that the replication factor in endoreplication cycles consisting of consecutive rounds of S phases without intervening mitosis could be more stable than that in the mitotic DNA replication. Furthermore, we have isolated the genomic and cDNA clones for DNA polymerase e and determined the whole primary structure. Immunohistochemical analyses indicates that DNA polymerase e expresses in all proliferative tissues during Drosophila development. Also, we isolated two null mutants by imprecise excision of the P element, with a 1.8 kb long deletion in the regulatory region from outside of the gene to just before TATA box. Both null mutants dies at late embryonic stages, indicating the DNA polymerase e is indispensable for Drosophila DNA replication. The heterozygotes for the null mutations also show a significant delay in larval and pupal growth. These phenotypes suggest that a loss of DNA polymerase e affects on DNA replication in mitotic as well as endoreplication tissues.
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[Publications] Lim, Y-M., Nishizawa, K., Nishi, Y., Tsuda, L., Inoue, Y.H.and Nishida, Y: "Genetic analysis of rolled that encodes a Drosophila MAP kinase."Genetics. 153. 763-771 (1999)
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[Publications] Yamaguchi, M., Hirose, F., Inoue, Y.H., Shiraki, M., Hayashi, Y., Nishi, Y.and Matsukage, A.: "Ectopic expression of human p53 inhibits entry into S phase and induces apoptosis in the Drosophila eye imaginal disc."Oncogene. 18. 6767-6775 (1999)
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[Publications] Hayashi, Y., Yamagishi, M., Nishimoto, Y., Taguchi, O., Matsukage, A.and Yamaguchi, M.: "A binding site for the transcription factor Grainyhead/Nuclear transcription factor-1 contributes to regulation of the Drosophila proliferating cell nuclear antigen gene promoter."J.Biol.Chem.. 274. 35080-35088 (1999)
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[Publications] Inoue, Y.H., do Carmo Avides, M., Shiraki, M., Deak, P., Yamaguchi, M., Nishimoto, Y., Matsukage, A.and Glover, D.M.: "Orbit, a novel microtubule-associated protein essential for mitosis in Drosophila."J.Cell Biol.. 149. 153-166 (2000)
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[Publications] Oshige, M., Yoshida, H., Hirose, F., Takata, K., Inoue, y.H., Aoyagi, N., Yamaguchi, M., Koiwai, O., Matsukage, A.and Sakaguchi, K.: "Molecular cloning and expression during development of the Drosophila gene for the catalytic subunit of DNA polymerase e."Gene. 256. 93-100 (2000)
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[Publications] Ohno, K., Takahashi, Y., Hirose, F., Inoue, Y.H., Taguchi, O., Nishida, Y., Matsukage, A.and Yamaguchi, M.: "Characterization of a Drosophila homologue of the human myelodysplasia/myeloid leukemia factor (MLF)."Gene. 260. 133-143 (2000)
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[Publications] Kwon, E.-J., Park, H.-S., Kim, Y.-S., Oh, E.-J., Nishida, Y., Matsukage, A., Yoo, M.-A.and Yamaguchi, M.: "Transcriptional regulation of the Drosophila raf proto-oncognen by drosophila STAT during development and immune response."J.Biol.Chem.. 275. 19824-19830 (2000)
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[Publications] Izumi, M., Yokoi, M., Nishikawa, N.S, Miyazawa, H., Sugino, A.Yamagishi, M., Yamaguchi, M., Matsukage, A., Yatagai, F.and Hanaoka, F.: "Transcription of the catalytic 180-kDa subunit gene of mouse DNA polymerase α is controlled by E2F, an Etsrelated transcription factor, and Sp1."Biochem.Biophys.Acta. 1492. 341-352 (2000)
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[Publications] Kwon, E.J., Oh, E.-J., Kim, Y.-S., Hirose, F., Ohno, K., Nishida, Y., Matsukage, A., Yamaguchi, M.and Yoo.M.-A.: "Transcriptional regulation of the Drosophila raf proto-oncogene by the transcription factor dE2F."Nucleic Acids Res.. (in press).
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