Karpova Nina, Bobinnec Yves, Fouix Sylvaine, Huitorel Philippe, Debec Alain
Laboratoire du Biologie du Développement, UMR7009 CNRS UPMC, Observatoire Océanologique, Villefranche-sur-mer, France.
Cell Motil Cytoskeleton. 2006 May;63(5):301-12. doi: 10.1002/cm.20124.
In this study we describe a novel Drosophila protein Jupiter, which shares properties with several structural microtubule-associated proteins (MAPs) including TAU, MAP2, MAP4. Jupiter is a soluble unfolded molecule with the high net positive charge, rich in Glycine. It possesses two degenerated repeats around the sequence PPGG, separated by a Serine-rich region. Jupiter associates with microtubules in vitro and, fused with the green fluorescent protein (GFP), is an excellent marker to follow microtubule dynamics in vivo. In a jupiter transgenic Drosophila strain generated by the "protein-trap" technique, Jupiter:GFP fusion protein localizes to the microtubule network through the cell cycle at the different stages of development. We found particularly high Jupiter:GFP concentrations in the young embryo, larval nervous system, precursors of eye photoreceptors and adult ovary. Moreover, from jupiter:gfp embryos we have established two permanent cell lines presenting strongly fluorescent microtubules during the whole cell cycle. In these cells, the distribution of the Jupiter:GFP fusion protein reproduces microtubule behavior upon treatment by the drugs colchicine and taxol. The jupiter cell lines and fly strain should be of wide interest for biologists interested in in vivo analysis of microtubule dynamics.
在本研究中，我们描述了一种新型果蝇蛋白朱庇特（Jupiter），它与包括TAU、MAP2、MAP4在内的几种结构性微管相关蛋白（MAPs）具有共同特性。朱庇特是一种具有高净正电荷、富含甘氨酸的可溶性未折叠分子。它在PPGG序列周围有两个退化重复序列，中间被一个富含丝氨酸的区域隔开。朱庇特在体外与微管结合，并且与绿色荧光蛋白（GFP）融合后，是追踪体内微管动态的优秀标记物。在通过“蛋白陷阱”技术产生的朱庇特转基因果蝇品系中，朱庇特：GFP融合蛋白在发育的不同阶段通过细胞周期定位于微管网络。我们在幼胚、幼虫神经系统、眼感光细胞前体和成年卵巢中发现了特别高的朱庇特：GFP浓度。此外，从朱庇特：gfp胚胎中我们建立了两个永久性细胞系，它们在整个细胞周期中都呈现出强烈荧光的微管。在这些细胞中，朱庇特：GFP融合蛋白的分布在秋水仙素和紫杉醇处理后重现了微管行为。朱庇特细胞系和果蝇品系对于对微管动态进行体内分析感兴趣的生物学家来说应该具有广泛的吸引力。
