Schäfer Gritt, Weber Susanne, Holz Anne, Bogdan Sven, Schumacher Sabine, Müller Arno, Renkawitz-Pohl Renate, Onel Susanne-Filiz
Fachbereich Biologie, Entwicklungsbiologie, Philipps-Universität Marburg, Karl-von-Frisch Str. 8, D-35043 Marburg, Germany.
Dev Biol. 2007 Apr 15;304(2):664-74. doi: 10.1016/j.ydbio.2007.01.015. Epub 2007 Jan 12.
In higher organisms, mononucleated myoblasts fuse to form multinucleated myotubes. During this process, myoblasts undergo specific changes in cell morphology and cytoarchitecture. Previously, we have shown that the actin regulator Kette (Hem-2/Nap-1) is essential for myoblast fusion. In this study, we describe the role of the evolutionary conserved Wiskott-Aldrich syndrome protein that serves as a regulator for the Arp2/3 complex for myoblast fusion. By screening an EMS mutagenesis collection, we discovered a new wasp allele that does not complete fusion during myogenesis. Interestingly, this new wasp3D3-035 allele is characterized by a disruption of fusion after precursor formation. The molecular lesion in this wasp allele leads to a stop codon preventing translation of the CA domain. Usually, the WASP protein exerts its function through the Arp2/3-interacting CA domain. Accordingly, a waspDeltaCA that is expressed in a wild-type background acts as dominant-negative during the fusion process. Furthermore, we show that the myoblast fusion phenotype of kette mutant embryos can be suppressed by reducing the gene dose of wasp3D3-035. Thus, Kette antagonizes WASP function during myoblast fusion.
在高等生物中,单核成肌细胞融合形成多核肌管。在此过程中,成肌细胞在细胞形态和细胞结构上会发生特定变化。此前,我们已经表明肌动蛋白调节因子Kette(Hem-2/Nap-1)对成肌细胞融合至关重要。在本研究中,我们描述了进化保守的威斯科特-奥尔德里奇综合征蛋白作为成肌细胞融合中Arp2/3复合物调节因子的作用。通过筛选一个EMS诱变文库,我们发现了一个新的wasp等位基因,其在肌生成过程中不能完成融合。有趣的是,这个新的wasp3D3-035等位基因的特征是在前体形成后融合中断。该wasp等位基因中的分子损伤导致一个终止密码子,阻止了CA结构域的翻译。通常情况下,WASP蛋白通过与Arp2/3相互作用的CA结构域发挥其功能。因此,在野生型背景中表达的waspDeltaCA在融合过程中起显性负作用。此外,我们表明通过降低wasp3D3-035的基因剂量可以抑制kette突变胚胎的成肌细胞融合表型。因此,Kette在成肌细胞融合过程中拮抗WASP的功能。