Rau A, Buttgereit D, Holz A, Fetter R, Doberstein S K, Paululat A, Staudt N, Skeath J, Michelson A M, Renkawitz-Pohl R
Developmental Biology, Philipps-Universität Marburg, 35032 Marburg, Germany.
Development. 2001 Dec;128(24):5061-73. doi: 10.1242/dev.128.24.5061.
Mutations in the rolling pebbles (rols) gene result in severe defects in myoblast fusion. Muscle precursor cells are correctly determined, but myogenesis does not progress significantly beyond this point because recognition and/or cell adhesion between muscle precursor cells and fusion-competent myoblasts is disturbed. Molecular analysis of the rols genomic region reveals two variant transcripts of rols due to different transcription initiation sites, rols6 and rols7. rols6 mRNA is detectable mainly in the endoderm during differentiation as well as in malpighian tubules and in the epidermis. By contrast, rols7 expression is restricted to the mesoderm and later to progenitor descendants during somatic and pharyngeal muscle development. Transcription starts at the extended germ band stage when progenitor/founder cells are determined and persists until stage 13. The proteins encoded by the rols gene are 1670 (Rols6) and 1900 (Rols7) amino acids in length. Both forms contain an N-terminal RING-finger motif, nine ankyrin repeats and a TPR repeat eventually overlaid by a coiled-coil domain. The longer protein, Rols7, is characterized by 309 unique N-terminal amino acids, while Rols6 is distinguishable by 79 N-terminal amino acids. Expression of rols7 in muscle founder cells indicates a function of Rols7 in these cells. Transplantation assays of rols mutant mesodermal cells into wild-type embryos show that Rols is required in muscle precursor cells and is essential to recruit fusion-competent myoblasts for myotube formation.
滚动卵石(rols)基因的突变会导致成肌细胞融合出现严重缺陷。肌肉前体细胞能够正确确定,但肌生成在此之后无法显著进展,因为肌肉前体细胞与具有融合能力的成肌细胞之间的识别和/或细胞黏附受到了干扰。对rols基因组区域的分子分析显示,由于转录起始位点不同,rols有两种可变转录本,即rols6和rols7。rols6 mRNA主要在分化过程中的内胚层以及马氏管和表皮中可检测到。相比之下,rols7的表达仅限于中胚层,在体细胞和咽肌发育过程中,随后仅限于祖细胞后代。转录在扩展胚带阶段开始,此时祖细胞/奠基细胞已确定,并持续到13阶段。rols基因编码的蛋白质长度分别为1670个氨基酸(Rols6)和1900个氨基酸(Rols7)。两种形式都包含一个N端的RING指基序、九个锚蛋白重复序列和一个TPR重复序列,最终被一个卷曲螺旋结构域覆盖。较长的蛋白质Rols7的特征是有309个独特的N端氨基酸,而Rols6则以79个N端氨基酸与之区分。rols7在肌肉奠基细胞中的表达表明Rols7在这些细胞中具有某种功能。将rols突变的中胚层细胞移植到野生型胚胎中的实验表明,Rols在肌肉前体细胞中是必需的,对于募集具有融合能力的成肌细胞形成肌管至关重要。