Molecular cytological studies on the relationship between mitochondrial genome damage and plasmodial senescence of the true slime mold, Physarum polycephalum

真粘菌多头绒泡菌线粒体基因组损伤与疟原虫衰老关系的分子细胞学研究

基本信息

  • 批准号:
    11440242
  • 负责人:
  • 金额:
    $ 5.95万
  • 依托单位:
  • 依托单位国家:
    日本
  • 项目类别:
    Grant-in-Aid for Scientific Research (B)
  • 财政年份:
    1999
  • 资助国家:
    日本
  • 起止时间:
    1999 至 2000
  • 项目状态:
    已结题

项目摘要

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.
我们发现从多头绒螯蟹中提取的mtDNA在弱离子溶液中会发生片段化。在STE(150 mM NaCl,TE)中观察到完整的mtDNA条带,但在TE(10 mM Tris-HCl,1 mM EDTA)和DW中出现了几条新的条带。在TE中,在67.7 kbp(α带)和5.0 kbp(β带)处出现两条离散带,而在DW中最终观察到至少17条离散带。对这些片段的定位表明,它们的切割位点是线粒体DNA的S1核酸酶敏感区。α和β片段来源于编码rRNA和ATP合酶(atpA)基因的区域。对幼年和老年疟原虫的mtDNA进行了等量的扩增,并测量了每个样品中β片段的频率。β带占总信号(包括背景)的比例在幼年疟原虫中估计为3.3 ~ 4.0%,而在老年疟原虫中增加到8.3 ~ 28.2%,α和β片段是体外DNA片段化的中间产物,在严酷的条件下(如DW)产生其余17个片段中的片段4、6、7和9。我们成功地测序了α和β片段的单链区域。在α片段的特定序列上至少有4个单链断裂。其中两个位于编码LrRNA的相对DNA链上的110 bp之间,另外两个位于编码atpA的相对DNA链上的86 bp之间。这两个区域的DNA裂解在体内破坏这两个基因,而片段4在体外破坏。直接检测这种单链断裂的引物延伸分析表明,他们的频率增加疟原虫衰老。随着年龄的增长,多头绒螯蟹线粒体DNA的单链断裂也会逐渐增多。

项目成果

期刊论文数量(70)
专著数量(0)
科研奖励数量(0)
会议论文数量(0)
专利数量(0)
河野重行: "ミトコンドリアの謎"講談社現代新書. 254 (1999)
河野茂之:《线粒体之谜》讲谈社现代新书 254 (1999)。
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    0
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  • 通讯作者:
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)
Kuriyama, H.、Takano, H.、Suzuki, L.、Uchida, H.、Kawano, S.、Kuroiwa, H. 和 Kuroiwa, H.:“编码含有锚蛋白重复序列​​的新型蛋白质的莱茵哈特合子特异性 cDNA 的表征
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    0
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Takahar,M.: "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)
Takahar,M.:“单细胞原始红藻 Cyanidium caldarium RK-1 中 ftsZ 基因的分离、表征和染色体定位。”Curr.Genet.. 37. 143-151 (2000)
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    0
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Sakai, A., Miyazawa, Y., Suzuki, T., Sasaki, N. Kawano, S. and Kuroiwa, T.: "Plastid gene expression during amyloplast formation in cultured tobacco cells."J. Plant Physiol.. 154. 71-78 (1999)
Sakai, A.、Miyazawa, Y.、Suzuki, T.、Sasaki, N. Kawano, S. 和 Kuroiwa, T.:“培养烟草细胞中淀粉体形成过程中的质体基因表达。”J.
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    0
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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)
Matsunaga, S.、Schutze, K.、Donnison, I.、Grant, S.、Kuroiwa, T. 和 Kawano, S.:“单一花粉分型与激光介导操作相结合”Plant J.. 20. 1-8
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KAWANO Shigeyuki其他文献

KAWANO Shigeyuki的其他文献

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{{ truncateString('KAWANO Shigeyuki', 18)}}的其他基金

An investigation into the plant dioecism by Correns' cross and the high throughput 4D screening
Correns 杂交和高通量 4D 筛选对植物雌雄异株的研究
  • 批准号:
    24657046
  • 财政年份:
    2012
  • 资助金额:
    $ 5.95万
  • 项目类别:
    Grant-in-Aid for Challenging Exploratory Research
Sex asymmetry and morphological structure deciding the direction of organellar inheritance in the algae
性别不对称和形态结构决定藻类细胞器遗传的方向
  • 批准号:
    22370024
  • 财政年份:
    2010
  • 资助金额:
    $ 5.95万
  • 项目类别:
    Grant-in-Aid for Scientific Research (B)
Silene latifolia and Microbotryum violaceum : Molecular, cellular and Morphological studies on plant sexuality and parasitic fungus operating it.
Silene latifolia 和 Microbotryum violaceum :植物性行为及其寄生真菌的分子、细胞和形态学研究。
  • 批准号:
    17370022
  • 财政年份:
    2005
  • 资助金额:
    $ 5.95万
  • 项目类别:
    Grant-in-Aid for Scientific Research (B)
Maternal inheritance of mitochondria : Molecular mechanisms for preferential digestion of male mtDNA
线粒体的母系遗传:优先消化男性线粒体DNA的分子机制
  • 批准号:
    15370027
  • 财政年份:
    2003
  • 资助金额:
    $ 5.95万
  • 项目类别:
    Grant-in-Aid for Scientific Research (B)
Molecular cytological studies on the mechanism of male mitochondria during maternal inheritance of mitochondria.
线粒体母系遗传过程中雄性线粒体机制的分子细胞学研究。
  • 批准号:
    13440246
  • 财政年份:
    2001
  • 资助金额:
    $ 5.95万
  • 项目类别:
    Grant-in-Aid for Scientific Research (B)

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    9032436
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    2014
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Immune Protection from Malaria: Age, Exposure Intensity, and the T Cell Response
疟疾的免疫保护:年龄、暴露强度和 T 细胞反应
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Insulin, IGF and Insulin Signaling: effects on Anopheles lifespan and immunity
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Insulin, IGF and Insulin Signaling: effects on Anopheles lifespan and immunity
胰岛素、IGF 和胰岛素信号传导:对按蚊寿命和免疫力的影响
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