Preferentially localized dynein and perinuclear dynactin associate with nuclear pore complex proteins to mediate genomic union during mammalian fertilization
Preferentially localized dynein and perinuclear dynactin associate with nuclear pore complex proteins to mediate genomic union during mammalian fertilization
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DOI：
10.1242/jcs.00784
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发表时间：
2003-12-01
影响因子：
4
通讯作者：
Schatten, G
中科院分区：
文献类型：
--
作者：
Payne, C;Rawe, V;Schatten, G
一旦亲本基因组结合，受精就完成了，并且需要在受精卵内的微管上将卵核迁移到精核（分别为雌性和雄性原核）。无论是原核结合微管的分子机制，也没有在调节原核运动的马达蛋白的作用已被充分的特点，和失败的合子发育在一些患者表明，它们有助于人类不育。基于女性原核迁移的负端方向，我们提出了细胞质动力蛋白和动力蛋白在与原核包膜和介导基因组结合中的作用。我们的研究结果表明，动力蛋白的中间和重链优先集中在女性原核周围，而动力蛋白亚基p150（胶合），p50和p62定位于两个原核的表面。抗动力蛋白和动力蛋白抗体的转染阻断了受精卵中雌性原核的迁移。孤雌激活卵母细胞和微管解聚受精卵显着减少本地化的动力蛋白的女性原核，但不抑制原核协会的动力蛋白。当从受精卵中免疫沉淀时，p150（Glued）与核孔复合物蛋白以及中间丝波形蛋白和动力蛋白结合。抗核孔蛋白和波形蛋白的抗体转染到受精卵中时抑制原核沉积。我们的结论是，优先本地化的动力蛋白和核周动力蛋白与核孔复合物和波形蛋白，并需要调解基因组的工会。这些数据表明，动力蛋白的积累和结合的女性原核精子紫菀微管，在那里它与dynactin，核孔蛋白和波形蛋白相互作用的模型。
Fertilization is complete once the parental genomes unite, and requires the migration of the egg nucleus to the sperm nucleus (female and male pronuclei, respectively) on microtubules within the inseminated egg. Neither the molecular mechanism of pronucleus binding to microtubules nor the role of motor proteins in regulating pronuclear motility has been fully characterized, and the failure of zygotic development in some patients suggests that they contribute to human infertility. Based on the minus-end direction of female pronuclear migration, we propose a role for cytoplasmic dynein and dynactin in associating with the pronuclear envelope and mediating genomic union. Our results show that dynein intermediate and heavy chains preferentially concentrate around the female pronucleus, whereas dynactin subunits p150(Glued), p50 and p62 localize to the surfaces of both pronuclei. Transfection of antibodies against dynein and dynactin block female pronuclear migration in zygotes. Both parthenogenetic activation in oocytes and microtubule depolymerization in zygotes significantly reduce the localization of dynein to the female pronucleus but do not inhibit the pronuclear association of dynactin. When immunoprecipitated from zygotes, p150(Glued) associates with nuclear pore complex proteins, as well as the intermediate filament vimentin and dynein. Antibodies against nucleoporins and vimentin inhibit pronuclear apposition when transfected into zygotes. We conclude that preferentially localized dynein and perinuclear dynactin associate with the nuclear pore complex and vimentin and are required to mediate genomic union. These data suggest a model in which dynein accumulates and binds to the female pronucleus on sperm aster microtubules, where it interacts with dynactin, nucleoporins and vimentin.