Donkor Fatima F, Mönnich Maren, Czirr Eva, Hollemann Thomas, Hoyer-Fender Sigrid
Göttinger Zentrum für Molekulare Biowissenschaften, Entwicklungsbiologie, Georg-August-Universität Göttingen, Justus-von-Liebig-Weg 11, 37077 Göttingen, Germany.
J Cell Sci. 2004 Sep 15;117(Pt 20):4643-51. doi: 10.1242/jcs.01303. Epub 2004 Aug 31.
Outer dense fibre protein 2 (ODF2) is a major protein of sperm tail outer dense fibres which are prominent sperm tail-specific cytoskeletal structures. Moreover, ODF2 was also identified as a widespread component of the centrosomal scaffold and was found to associate preferentially with the appendages of the mother centriole [Nakagawa, Y., Yamane, Y., Okanoue, T., Tsukita, S. and Tsukita, S. (2001) Mol. Biol. Cell 12, 1687-1697]. Secondary structure predictions indicated ODF2 as an overall coiled-coil protein with a putative fibre forming capacity. To investigate its potential functions in generating the centrosomal scaffold and in microtubule nucleation we asked whether ODF2 is able to form a fibrillar structure by self-association in vivo and if it interacts with microtubules. By cytological investigation of transfected mammalian cells expressing ODF2-GFP fusion proteins and in vitro coprecipitation assays we could demonstrate that ODF2 is a self-interacting protein that forms a fibrillar structure partially linked to the microtubule network. Microtubule cosedimentation and coprecipitation assays indicated ODF2 as a microtubule-associated protein. However, we could not demonstrate a direct interaction of ODF2 with tubulin, suggesting that binding of endogenous ODF2 to the axonemal as well as to centrosomal microtubules may be mediated by, as yet, unknown proteins.
外致密纤维蛋白2(ODF2)是精子尾部外致密纤维的主要蛋白质,外致密纤维是精子尾部特有的突出细胞骨架结构。此外,ODF2还被鉴定为中心体支架的广泛组成成分,并被发现优先与母中心粒的附属物相关联[中川洋、山根洋、冈上彻、筑田司和筑田节子(2001年)《分子生物学细胞》第12卷,第1687 - 1697页]。二级结构预测表明ODF2是一种整体呈卷曲螺旋结构的蛋白质,具有假定的纤维形成能力。为了研究其在生成中心体支架和微管成核中的潜在功能,我们探究了ODF2是否能够在体内通过自组装形成纤维状结构,以及它是否与微管相互作用。通过对表达ODF2 - GFP融合蛋白的转染哺乳动物细胞进行细胞学研究以及体外共沉淀分析,我们能够证明ODF2是一种自我相互作用的蛋白质,它形成了一种部分与微管网络相连的纤维状结构。微管共沉降和共沉淀分析表明ODF2是一种微管相关蛋白。然而,我们无法证明ODF2与微管蛋白之间存在直接相互作用,这表明内源性ODF2与轴丝以及中心体微管的结合可能是由尚未知晓的蛋白质介导的。