Willis John H, Munro Edwin, Lyczak Rebecca, Bowerman Bruce
Institute of Molecular Biology, University of Oregon, Eugene, OR 97403, USA.
Mol Biol Cell. 2006 Mar;17(3):1051-64. doi: 10.1091/mbc.e05-09-0886. Epub 2006 Jan 11.
Animal genomes each encode multiple highly conserved actin isoforms that polymerize to form the microfilament cytoskeleton. Previous studies of vertebrates and invertebrates have shown that many actin isoforms are restricted to either nonmuscle (cytoplasmic) functions, or to myofibril force generation in muscle cells. We have identified two temperature-sensitive and semidominant embryonic-lethal Caenorhabditis elegans mutants, each with a single mis-sense mutation in act-2, one of five C. elegans genes that encode actin isoforms. These mutations alter conserved and adjacent amino acids predicted to form part of the ATP binding pocket of actin. At the restrictive temperature, both mutations resulted in aberrant distributions of cortical microfilaments associated with abnormal and striking membrane ingressions and protrusions. In contrast to the defects caused by these dominant mis-sense mutations, an act-2 deletion did not result in early embryonic cell division defects, suggesting that additional and redundant actin isoforms are involved. Accordingly, we found that two additional actin isoforms, act-1 and act-3, were required redundantly with act-2 for cytoplasmic function in early embryonic cells. The act-1 and -3 genes also have been implicated previously in muscle function. We found that an ACT-2::GFP reporter was expressed cytoplasmically in embryonic cells and also was incorporated into contractile filaments in adult muscle cells. Furthermore, one of the dominant act-2 mutations resulted in uncoordinated adult movement. We conclude that redundant C. elegans actin isoforms function in both muscle and nonmuscle contractile processes.
动物基因组各自编码多种高度保守的肌动蛋白异构体,这些异构体聚合形成微丝细胞骨架。先前对脊椎动物和无脊椎动物的研究表明,许多肌动蛋白异构体要么局限于非肌肉(细胞质)功能,要么局限于肌肉细胞中肌原纤维的力产生。我们鉴定出两种温度敏感且半显性胚胎致死的秀丽隐杆线虫突变体,每个突变体在act-2基因中都有一个错义突变,act-2是秀丽隐杆线虫五个编码肌动蛋白异构体的基因之一。这些突变改变了预测形成肌动蛋白ATP结合口袋一部分的保守且相邻的氨基酸。在限制温度下,这两种突变都导致皮质微丝的异常分布,伴有异常且显著的膜内陷和突出。与这些显性错义突变导致的缺陷相反,act-2基因缺失并未导致早期胚胎细胞分裂缺陷,这表明还有其他冗余的肌动蛋白异构体参与其中。因此,我们发现另外两种肌动蛋白异构体act-1和act-3与act-2在早期胚胎细胞的细胞质功能中发挥冗余作用。act-1和act-3基因先前也被认为与肌肉功能有关。我们发现一个ACT-2::GFP报告基因在胚胎细胞的细胞质中表达,并且也被整合到成年肌肉细胞的收缩丝中。此外,一个显性act-2突变导致成年个体运动不协调。我们得出结论,秀丽隐杆线虫的冗余肌动蛋白异构体在肌肉和非肌肉收缩过程中均发挥作用。