Abstract
Null alpha-actinin mutations in Drosophila are lethal and produce conspicuous defects in muscle structure and function. Here, we used transgene rescue to examine the requirements for alpha-actinin function in vivo. First, we tested the ability of a cDNA-based transgene encoding the adult muscle isoform of alpha-actinin under control of the heterologous ubiquitin promoter to rescue the lethality of null alpha-actinin mutations. Successful rescue indicated that alternative splicing, which also generates larval muscle and non-muscle isoforms, was not essential for viability and that there were no strict spatial or temporal requirements for alpha-actinin expression. Secondly, chimeric transgenes, with functional domains of alpha-actinin replaced by similar domains from spectrin, were tested for their ability to rescue alpha-actinin mutants. Replacement of either the actin binding domain or the EF hand calcium binding domain yielded inactive proteins, indicating that these conserved domains were not functionally equivalent. Thirdly, the length of alpha-actinin was modified by adding a 114 amino acid structural repeat from alpha-spectrin to the center of the rod domain of alpha-actinin. Addition of this sequence module was expected to increase the length of the native alpha-actinin molecule by at least 15%. yet was fully compatible with alpha-actinin function as measured by rescued lethality and flight. Thus, unexpectedly, the exact length of alpha-actinin was not critical to its function in the muscle Z disk.
