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在海胆胚胎的有丝分裂和纤毛发生周期中,驱动蛋白-II亚基KAP从纤毛到细胞核的重新分布。

Redistribution of the kinesin-II subunit KAP from cilia to nuclei during the mitotic and ciliogenic cycles in sea urchin embryos.

作者信息

Morris Robert L, English Christopher N, Lou Julia E, Dufort Fay J, Nordberg Joshua, Terasaki Mark, Hinkle Beth

机构信息

Department of Biology, Wheaton College, Norton, MA 02766, USA.

出版信息

Dev Biol. 2004 Oct 1;274(1):56-69. doi: 10.1016/j.ydbio.2004.06.017.

Abstract

KAP is the non-motor subunit of the heteromeric plus-end directed microtubule (MT) motor protein kinesin-II essential for normal cilia formation. Studies in Chlamydomonas have demonstrated that kinesin-II drives the anterograde intraflagellar transport (IFT) of protein complexes along ciliary axonemes. We used a green fluorescent protein (GFP) chimera of KAP, KAP-GFP, to monitor movements of this kinesin-II subunit in cells of sea urchin blastulae where cilia are retracted and rebuilt with each mitosis. As expected if involved in IFT, KAP-GFP localized to apical cytoplasm, basal bodies, and cilia and became concentrated on basal bodies of newly forming cilia. Surprisingly, after ciliary retraction early in mitosis, KAP-GFP moved into nuclei before nuclear envelope breakdown, was again present in nuclei after nuclear envelope reformation, and only decreased in nuclei as ciliogenesis reinitiated. Nuclear transport of KAP-GFP could be due to a putative nuclear localization signal and nuclear export signals identified in the sea urchin KAP primary sequence. Our observation of a protein involved in IFT being imported into the nucleus after ciliary retraction and again after nuclear envelope reformation suggests KAP115 may serve as a signal to the nucleus to reinitiate cilia formation during sea urchin development.

摘要

KAP是异源多聚体正向微管(MT)驱动蛋白驱动蛋白-II的非运动亚基,对正常纤毛形成至关重要。衣藻研究表明,驱动蛋白-II沿着纤毛轴丝驱动蛋白质复合物的顺行鞭毛内运输(IFT)。我们使用KAP的绿色荧光蛋白(GFP)嵌合体KAP-GFP来监测这种驱动蛋白-II亚基在海胆囊胚细胞中的运动,在海胆囊胚细胞中,纤毛在每次有丝分裂时都会缩回并重新形成。如果参与IFT,KAP-GFP预期会定位于顶端细胞质、基体和纤毛,并在新形成纤毛的基体上富集。令人惊讶的是,在有丝分裂早期纤毛缩回后,KAP-GFP在核膜破裂前进入细胞核,在核膜重新形成后再次出现在细胞核中,并且仅在纤毛发生重新启动时在细胞核中减少。KAP-GFP的核运输可能归因于在海胆KAP一级序列中鉴定出的假定核定位信号和核输出信号。我们观察到一种参与IFT的蛋白质在纤毛缩回后以及核膜重新形成后再次被导入细胞核,这表明KAP115可能作为一种信号告知细胞核在海胆发育过程中重新启动纤毛形成。

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