Fischer Janice A, Acosta Shelley, Kenny Andrew, Cater Courtney, Robinson Christina, Hook Jay
Section of Molecular Cell and Developmental Biology, Institute for Cellular and Molecular Biology, University of Texas, Austin, Texas 78712, USA.
Genetics. 2004 Nov;168(3):1385-93. doi: 10.1534/genetics.104.028662.
The Drosophila klarsicht (klar) gene is required for developmentally regulated migrations of photoreceptor cell nuclei in the eye. klar encodes a large ( approximately 250 kD) protein with only one recognizable amino acid sequence motif, a KASH (Klar, Anc-1, Syne-1 homology) domain, at its C terminus. It has been proposed that Klar facilitates nuclear migration by linking the nucleus to the microtubule organizing center (MTOC). Here we perform genetic and immunohistochemical experiments that provide a critical test of this model. We analyze mutants in the endogenous klar gene and also flies that express deleted forms of Klar protein from transgenes. We find that the KASH domain of Klar is critical for perinuclear localization and for function. In addition, we find that the N-terminal portion of Klar is also important for function and contains a domain that localizes the protein to microtubules apical to the nucleus. These results provide strong support for a model in which Klar links the nucleus to the MTOC.
果蝇的klar基因是眼内光感受器细胞核发育调控迁移所必需的。klar编码一种大约250 kD的大蛋白,其C端只有一个可识别的氨基酸序列基序,即KASH(Klar、Anc-1、Syne-1同源性)结构域。有人提出,Klar通过将细胞核与微管组织中心(MTOC)相连来促进核迁移。在此,我们进行了遗传学和免疫组织化学实验,对该模型进行了关键检验。我们分析了内源性klar基因的突变体以及从转基因中表达缺失形式Klar蛋白的果蝇。我们发现,Klar的KASH结构域对于核周定位和功能至关重要。此外,我们发现Klar的N端部分对于功能也很重要,并且包含一个将该蛋白定位于细胞核顶端微管的结构域。这些结果为Klar将细胞核与MTOC相连的模型提供了有力支持。