Stronach B E, Siegrist S E, Beckerle M C
Department of Biology, University of Utah, Salt Lake City 84112, USA.
J Cell Biol. 1996 Sep;134(5):1179-95. doi: 10.1083/jcb.134.5.1179.
The LIM domain defines a zinc-binding motif found in a growing number of eukaryotic proteins that regulate cell growth and differentiation during development. Members of the cysteine-rich protein (CRP) family of LIM proteins have been implicated in muscle differentiation in vertebrates. Here we report the identification and characterization of cDNA clones encoding two members of the CRP family in Drosophila, referred to as muscle LIM proteins (Mlp). Mlp60A encodes a protein with a single LIM domain linked to a glycine-rich region. Mlp84B encodes a protein with five tandem LIM-glycine modules. In the embryo, Mlp gene expression is spatially restricted to somatic, visceral, and pharyngeal muscles. Within the somatic musculature, Mlp84B transcripts are enriched at the terminal ends of muscle fibers, whereas Mlp60A transcripts are found throughout the muscle fibers. The distributions of the Mlp60A and Mlp84B proteins mirror their respective mRNA localizations, with Mlp84B enrichment occurring at sites of muscle attachment. Northern blot analysis revealed that Mlp gene expression is developmentally regulated, showing a biphasic pattern over the course of the Drosophila life cycle. Peaks of expression occur late in embryogenesis and during metamorphosis, when the musculature is differentiating. Drosophila Mlp60A and Mlp84B, like vertebrate members of the CRP family, have the ability to associate with the actin cytoskeleton when expressed in rat fibroblast cells. The temporal expression and spatial distribution of muscle LIM proteins in Drosophila are consistent with a role for Mlps in myogenesis, late in the differentiation pathway.
LIM结构域定义了一种锌结合基序,该基序存在于越来越多的真核生物蛋白质中,这些蛋白质在发育过程中调节细胞生长和分化。富含半胱氨酸蛋白(CRP)家族的LIM蛋白成员与脊椎动物的肌肉分化有关。在此,我们报告了果蝇中编码CRP家族两个成员的cDNA克隆的鉴定和特征,这两个成员被称为肌肉LIM蛋白(Mlp)。Mlp60A编码一种具有单个LIM结构域并与富含甘氨酸区域相连的蛋白质。Mlp84B编码一种具有五个串联LIM-甘氨酸模块的蛋白质。在胚胎中,Mlp基因表达在空间上局限于体壁、内脏和咽肌。在体壁肌肉组织中,Mlp84B转录本在肌纤维末端富集,而Mlp60A转录本则遍布整个肌纤维。Mlp60A和Mlp84B蛋白的分布反映了它们各自mRNA的定位,Mlp84B在肌肉附着部位富集。Northern印迹分析表明,Mlp基因表达受发育调控,在果蝇生命周期中呈现双相模式。表达高峰出现在胚胎发育后期和变态期间,此时肌肉组织正在分化。果蝇的Mlp60A和Mlp84B与CRP家族的脊椎动物成员一样,当在大鼠成纤维细胞中表达时,具有与肌动蛋白细胞骨架结合的能力。果蝇中肌肉LIM蛋白的时间表达和空间分布与Mlp在分化途径后期的肌生成中的作用一致。