Clemente Giuliana D, Hannaford Matthew R, Beati Hamze, Kapp Katja, Januschke Jens, Griffis Eric R, Müller Hans-Arno J
Division of Cell and Developmental Biology, School of Life Sciences, University of Dundee, Dundee DD1 5EH, UK.
Institut für Biologie, Fachgebiet Entwicklungsgenetik, Universität Kassel, Heinrich-Plett Str. 40, 34321 Kassel, Germany.
J Dev Biol. 2018 Mar 30;6(2):9. doi: 10.3390/jdb6020009.
Spindly was originally identified as a specific regulator of Dynein activity at the kinetochore. In early prometaphase, Spindly recruits the Dynein/Dynactin complex, promoting the establishment of stable kinetochore-microtubule interactions and progression into anaphase. While details of Spindly function in mitosis have been worked out in cultured human cells and in the zygote, the function of Spindly within the context of an organism has not yet been addressed. Here, we present loss- and gain-of-function studies of Spindly using transgenic RNAi in . Knock-down of Spindly in the female germ line results in mitotic arrest during embryonic cleavage divisions. We investigated the requirements of Spindly protein domains for its localisation and function, and found that the carboxy-terminal region controls Spindly localisation in a cell-type specific manner. Overexpression of Spindly in the female germ line is embryonic lethal and results in altered egg morphology. To determine whether Spindly plays a role in post-mitotic cells, we altered Spindly protein levels in migrating cells and found that ovarian border cell migration is sensitive to the levels of Spindly protein. Our study uncovers novel functions of Spindly and a differential, functional requirement for its carboxy-terminal region in .
Spindly最初被鉴定为动粒处动力蛋白活性的特定调节因子。在早前期，Spindly招募动力蛋白/动力蛋白激活蛋白复合体，促进稳定的动粒-微管相互作用的建立并推动进入后期。虽然Spindly在有丝分裂中的功能细节已在培养的人类细胞和受精卵中得到阐明，但Spindly在生物体中的功能尚未得到研究。在此，我们利用转基因RNA干扰技术对Spindly进行了功能缺失和功能获得性研究。在雌性生殖系中敲低Spindly会导致胚胎卵裂期的有丝分裂停滞。我们研究了Spindly蛋白结构域对其定位和功能的需求，发现羧基末端区域以细胞类型特异性方式控制Spindly的定位。在雌性生殖系中过表达Spindly会导致胚胎致死并使卵形态改变。为了确定Spindly是否在有丝分裂后细胞中发挥作用，我们改变了迁移细胞中Spindly蛋白的水平，发现卵巢边缘细胞迁移对Spindly蛋白水平敏感。我们的研究揭示了Spindly的新功能及其羧基末端区域在[具体生物]中的不同功能需求。
