The stereotyped striation of myofibrils is a conserved feature of muscle organization that is critical to its function. Although most components that constitute the basic myofibrils are well-characterized biochemically and are conserved across the animal kingdom, the mechanisms leading to the precise assembly of sarcomeres, the basic units of myofibrils, are poorly understood. To gain insights into this process, we investigated the functional relationships of sarcomeric protein complexes. Specifically, we systematically analyzed, using either RNAi in primary muscle cells or available genetic mutations, the organization of myofibrils in Drosophila muscles that lack one or more sarcomeric proteins. Our study reveals that the thin and thick filaments are mutually dependent on each other for striation. Further, the tension sensor complex comprised of zipper/Zasp/α-actinin is involved in stabilizing the sarcomere but not in its initial formation. Finally, integrins appear essential for the interdigitation of thin and thick filaments that occurs prior to striation. Thus, sarcomere formation occurs by the coordinated assembly of multiple latent protein complexes, as opposed to sequential assembly.
Muscle functionality relies on the correct assembly of myofibrils, which are composed of tandem arrays of basic functional contractile units called the sarcomeres. Many mutations in genes encoding sarcomeric proteins cause muscle diseases such as congenital myopathy and dilated cardiac hypertrophy. Understanding the process of sarcomere assembly is not only relevant to the understanding of how protein complexes interact to form complex supra-molecular structures, but also of great significance to medicine for muscle diseases. Here, by taking advantage of our newly developed primary muscle cell culture method, we reevaluate sarcomere assembly by systematically analyzing the functional relationship of sarcomeric proteins using RNA interference or genetic ablation techniques. Our analysis leads us to propose a “two-state” model whereby sarcomeric proteins exist either in the “chaotic” state with independently assembled differential functional complexes or the “highly ordered suprastructure” state made from these complexes. Because we fail to detect any previously hypothesized sarcomere assembly intermediates in our system, our data support the model that sarcomere assembly is a highly coordinated process mediated by multiple latent protein complexes and does not occur in a step-wise fashion.