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
• 负责人: KAWANO Shigeyuki
• 金额: $ 8.7万
• 依托单位: The University of Tokyo
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for Scientific Research (B)
• 财政年份: 2001
• 资助国家: 日本
• 起止时间: 2001 至 2002
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/en/grant/KAKENHI-PROJECT-13440246/
• 关键词: maternal inheritance; Physarum polycephalum; Silene latifolia; mitochondria; mtDNA; matA hierarchy; selective digestion; DNase

The fluorescence of mtDNA stained with DAPI in half of the mitochondria decreased simultaneously to give small spots, and then disappeared completely about 1 h after nuclear fusion, while the other mt-nucleoids and all of the mitochondrial sheaths remained unchanged in Physarum polycehalum. PCR analysis of single zygote cells confirmed that the loss was limited to mtDNA from one parent. The mt-nucleoid protein also dispersed soon after the digestion of mtDNA. The dispersed mt-nucleoid protein was eliminated from mitochondria. The vacant mitochondrial sheaths were gradually eliminated by 60 h after mating. Using six mating types, the transmission patterns of mtDNA were examined in all possible crosses. In 39 of 60 crosses, strict uniparental inheritance was confirmed in accordance with a hierarchy of relative sexuality. In the other crosses, however, mtDNA from both parents was transmitted to plasmodia. The ratio of parental mtDNA was estimated to be from 1:1 to 1:10^<-4>. Nevertheless, the matA hierarchy was followed. In these crosses, the mtDNA was incompletely digested, and mtDNA replicated during subsequent plasmodial development. We conclude that the rapid, selective digestion of mtDNA promotes the uniparental inheritance of mitochondria ; when this fails, biparental inheritance occurs. We used DNA zymography assays with native PAGE and SDS-PAGE to investigate the DNase activities in young zygotes, and identified two novel mitochondrial DNase activities a 50 kDa Ca^<2+>-dependent DNase, and a 13-kDa Mn^<2+>-+dependent DNase. We propose that the regulation of the mitochondrial nuclease by some unknown factor is the key to digestion of the mtDNA from one parent.

用DAPI染色的MtDNA的荧光同时减少以产生小斑点，然后在核融合后大约1小时完全消失，而其他MT核苷酸和所有线粒体鞘层在Pholyarum Polycehalum中保持不变。对单个合子细胞的PCR分析证实，损失仅限于一个父母的mtDNA。 MtDNA消化后不久，MT核苷蛋白也分散。从线粒体中消除了分散的MT核苷蛋白。交配后60小时，逐渐消除空缺的线粒体鞘。使用六种交配类型，在所有可能的杂交中检查了mtDNA的传输模式。在60个十字架中的39个中，根据相对性的层次结构确认了严格的单亲继承。然而，在其他十字架中，父母双方的mtDNA被传播到疟原虫。父母mtDNA的比率估计为1：1至1:10^<-4>。尽管如此，遵循了MATA等级制度。在这些十字架中，mtDNA被不完全消化，并在随后的静态发育过程中复制了mtDNA。我们得出的结论是，mtDNA的快速，选择性消化促进了线粒体的单亲遗传。当这种情况失败时，两国继承就会发生。我们使用了具有天然页面和SDS-PAGE的DNA zymography测定法研究了年轻的Zygotes中的DNase活性，并鉴定了两种新型的线粒体DNase活性A A 50 kDa ca^<2+> - 依赖性DNase和13 kDa Mn^<2+> - +++++依赖性的DNase。我们建议通过某个未知因子对线粒体核酸酶调节是从一个父母那里消化mtDNA的关键。

项目成果

Takano, H., Abe, T., Sakurai, R., Kawano, S., Sasaki, N., Kuroiwa, T.: "The complete DNA sequence of the mitochondrial genome of Physarum polycephalum"Mol.Gen.Genet.. 264. 539-545 (2001)

Takano, H.、Abe, T.、Sakurai, R.、Kawano, S.、Sasaki, N.、Kuroiwa, T.：“多头绒泡菌线粒体基因组的完整 DNA 序列”Mol.Gen.Genet..

Moriyama, Y., Kawano, S.: "Rapid, selective digestion of mitochondrial DNA in accordance with the matA hierarchy of multiallelic mating-types in the mitochondrial inheritance of Physarum polycephalum."Genetics. 164. 963-975 (2003)

Moriyama, Y., Kawano, S.：“根据多头绒泡菌线粒体遗传中多等位基因交配类型的 matA 层次结构，对线粒体 DNA 进行快速、选择性消化。”遗传学。

Uchida, W., Matsunaga, S., Sugiyama, R., Kazama, Y., Kawano, S.: "Morphological development of anthers induced by the dimorphic smut fungus Microbotryum violaceum in female flowers of the dioecious plant Silene latifolia."Plant. 218. 240-248 (2003)

Uchida, W.、Matsunaga, S.、Sugiyama, R.、Kazama, Y.、Kawano, S.：“雌雄异株植物 Silene latifolia 雌花中二相黑粉病真菌 Microbotryum violaceum 诱导花药的形态发育。”

Sasaki, N., Kuroiwa, H., Nishitani, C., Takano, H., Higashiyama, T., Kobayashi, T., Shirai, Y., Sakai, S., Kawano, S., Murakami-Murofushi, K., Kuroiwa, T.: "Glom is a novel mitochondrial DNA packaging protein in Physarum polycephalum and causes intense ch

佐佐木，N.，黑岩，H.，西谷，C.，高野，H.，东山，T.，小林，T.，白井，Y.，酒井，S.，河野，S.，村上室伏，K

森山陽介: "植物工学別冊17、ミトコンドリアの動態と遺伝"秀潤社. 130-138 (2002)

森山洋介：《植物工程特刊第17卷，线粒体动力学与遗传学》书俊社130-138（2002）。