2001 Fiscal Year Final Research Report Summary
Molecular cytological studies on the relationship between mitochondrial genome damage and plasmodial senescence of the true slime mold, Physarum polycephalum
| Project/Area Number |
11440242
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| Research Category |
Grant-in-Aid for Scientific Research (B)
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| Allocation Type | Single-year Grants |
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| Section | 一般 |
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| Research Field |
生物形態・構造
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| Research Institution | The University of Tokyo |
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Principal Investigator |
KAWANO Shigeyuki Graduate School of Sciences, The University of Tokyo, Professor, 大学院・新領域創成科学研究科, 教授 (70161338)
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| Co-Investigator(Kenkyū-buntansha) |
TAKANO Hiroyoshi Faculty of Science, Kumamoto University, Associate Professor, 理学部, 助教授 (70242104)
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| Project Period (FY) |
1999 – 2000
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| Keywords | Physarum polycephalum / plasmodium / aging / senescence / mitochondria / single-stranded breakage / dysfunction / genome damage |
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| Research Abstract |
We found that mtDNA isolated from P. polycephalum fragmented itself in weak ionic solutions. The intact mtDNA band was seen in STE (150 mM NaCl, TE), but several novel bands appeared in TE (10 mM Tris-HC1, 1 mM EDTA) and DW. In TE, two discrete bands appeared at 67.7 kbp (α band) and 5.0 kbp (β band), whereas at least 17 discrete bands were finally observed in DW. Mapping these fragments demonstrated that their cleavage sites are S1 nuclease sensitive regions of the mtDNA. The α and β fragments are derived from the region encoding rRNAs and the ATP synthase (atpA) gene. Equal amounts of mtDNA from juvenile and aged plasmodia were electrophoresed and the frequency of the β fragment in each sample was measured. The ratio of the β band to the total signal including background was estimated to be 3.3 to 4.0 % in juvenile plasmodia whereas it increased to 8.3 to 28.2 % in aged plasmodia.
The α and β fragments are intermediates of in vitro DNA fragmentation and yield fragments 4, 6, 7 and 9 of the other final 17 fragments under severe condition such as DW. We succeeded to sequence the single stranded regions of the α and β fragments. There are at least 4 single stranded breakages on specific sequences of the α fragment. Two of them are located between 110 bp on opposite DNA strands coding LrRNA, and the other two are located between 86 bp on opposite DNA strands coding atpA. The DNA cleavages of these two regions disrupt the two genes in vivo while the fragment 4 in vitro. Direct detection of such single stranded breakages by a primer-extension analysis showed that the frequency of them increased as plasmodial senescence. The single-stranded breakage of mtDNA of P. polycephalum may accumulate with age.
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[Publications] Matsunaga, S., Kawano, S., Michimoto, T., Higashiyama, T., Nakao, S. and Kuroiwa, T.: "Semi-automatic laser beam microdissection of the Y chromosome and analysis of Y chromosome DNA in a dioecious plant, Silene latifolia."Plant Cell Physiol.. 40. 60-68 (1999)
Description
「研究成果報告書概要(欧文)」より
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[Publications] Kuriyama, H., Takano, H., Suzuki, L., Uchida, H., Kawano, S., Kuroiwa, H. and Kuroiwa, H.: "Characterization of reinhardtii zygote-specific cDNAs that encode novel proteins containing ankyrin repeats and WW domains."Plant Physiol.. 119. 873-884 (1999)
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「研究成果報告書概要(欧文)」より
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[Publications] Miyazawa, Y., Sakai, A., Miyagishima, S., Takano, T., Kawano, S. and Kuroiwa, T.: "Auxin and cytokinin have opposite effects on amyloplast development and the expression of starch synthesis genes in\ cultured Bright Yellow-2 tobacco cell."Plant Physiol.. 121. 461-470 (1999)
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[Publications] Takahara, M., Takahashi, H., Matsunaga, S., Sakai, A., Kawano, S. and Kuroiwa, T.: "Two types of ftsZ genes isolated from the unicellular primitive red alga Galdieria sulphuraria."Plant Cell Physiol.. 40. 784-791 (1999)
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「研究成果報告書概要(欧文)」より
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[Publications] Uchida, H., Suzuki, L., Anai, T., Doi, K., Takano, H., Yamashita, H., Oka, T., Kawano, S., Tomizawa, K., Kawazu, T., Kuroiwa, H. and Kuroiwa,T.: "A pair of invertedly repeated genes in Chlamydomonas reinhardtii encodes a zygote-specific protein whose expression is UV-sensitive."Curr. Genet.. 36. 232-240 (1999)
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「研究成果報告書概要(欧文)」より
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[Publications] Matsunaga, S., Schutze, K., Donnison, I., Grant, S., Kuroiwa, T. and Kawano, S.: "Single pollen typing combined with laser-mediated manipulation"Plant J.. 20. 1-8 (1999)
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[Publications] Misumi, O., Suzuki, L., Nishimura, Y., Sakai, A., Kawano, S., Kuroiwa, H. and Kuroiwa, T.: "Isolation and phenotypic characterization of Chlamydomonas reinhardtii mutants defective in chloroplast DNA segregation"Protoplasma. 209. 273-282 (1999)
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「研究成果報告書概要(欧文)」より
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[Publications] Takahara, M., Tahahashi, H., Matsunaga, S., Sakai, A., Kawano, S. and Kuroiwa, T.: "Isolation, charachterization, and chromosomal mapping of an ftsZ gene from the unicellular primitive red alga Cyanidium caldarium RK-1"Curr. Genet.. 37. 143-151 (2000)
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「研究成果報告書概要(欧文)」より
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[Publications] Takano, H., Abe, T., Sakurai, R., Kawano, S., Sasaki, N. and Kuroiwa, T.: "The complete DNA sequence of the mitochondrial genome of Physarum polycephalum"Mol. Gen. Genet.. 264. 539-545 (2001)
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「研究成果報告書概要(欧文)」より
