Paoletti A, Moudjou M, Paintrand M, Salisbury J L, Bornens M
Institut Curie, Section Recherche, UMR 144 du CNRS, Paris, France.
J Cell Sci. 1996 Dec;109 ( Pt 13):3089-102. doi: 10.1242/jcs.109.13.3089.
Centrin is a member of the calcium-binding EF-hand protein superfamily present in centrosomes of widely divergent species. Investigating the cellular distribution of human centrin by both immunofluorescence and cell fractionation, we report that centrin is biochemically complex in human cells, displaying as much as ten isoforms in 2-D electrophoresis. This suggests that centrin may be subject to multiple regulations. Strikingly, more than 90% of centrin is not associated with the centrosome fraction. The centrosome-associated centrin, however, displays a specific pattern in 2-D electrophoresis and is concentrated within the distal lumen of the centrioles, where a complex structure has been previously described. This precise localization allows the resolution of centrioles at the optical level throughout the cell cycle and provides a valuable tool for monitoring centriole duplication. To get insights on centrin function, we performed injection experiments of recombinant heterologous centrin in two-cell stage frog embryos in an attempt to produce dominant negative effects. We report that green algae and human centrin delay cleavage and promote the formation of abnormal blastomeres in which the distribution of microtubule asters and of nuclei is dramatically impaired. This suggests that centrin could be involved in the centrosome reproduction cycle, in the coordination of cytoplasmic and nuclear division or in cytokinesis.
中心蛋白是一种存在于广泛不同物种中心体中的钙结合EF手蛋白超家族成员。通过免疫荧光和细胞分级分离研究人类中心蛋白的细胞分布,我们发现中心蛋白在人类细胞中具有生化复杂性,在二维电泳中显示多达十种同工型。这表明中心蛋白可能受到多种调控。令人惊讶的是,超过90%的中心蛋白与中心体部分不相关。然而,与中心体相关的中心蛋白在二维电泳中呈现出特定模式,并集中在中心粒的远端腔内,此前已描述过那里有复杂结构。这种精确的定位使得在整个细胞周期中都能在光学水平上分辨中心粒,并为监测中心粒复制提供了有价值的工具。为了深入了解中心蛋白的功能,我们在两细胞期青蛙胚胎中进行了重组异源中心蛋白的注射实验,试图产生显性负效应。我们发现绿藻和人类中心蛋白会延迟卵裂并促进异常卵裂球的形成,其中微管星状体和细胞核的分布受到显著损害。这表明中心蛋白可能参与中心体复制周期、细胞质和核分裂的协调或胞质分裂。