caa a22 4500 475744950 CHVBK 20180406123510.0 cr unu---uuuuu 170329e20001001xx s 000 0 eng 10.1023/A:1005699608893 doi (NATIONALLICENCE)springer-10.1023/A:1005699608893 Genetic analysis of the requirements for α-actinin function [Elektronische Daten] [Ronald Dubreuil, Ping Wang] Null α-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 α-actinin function in vivo. First, we tested the ability of a cDNA-based transgene encoding the adult muscle isoform of α-actinin under control of the heterologous ubiquitin promoter to rescue the lethality of null α-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 α-actinin expression. Secondly, chimeric transgenes, with functional domains of α-actinin replaced by similar domains from spectrin, were tested for their ability to rescue α-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 α-actinin was modified by adding a 114 amino acid structural repeat from α-spectrin to the center of the rod domain of α-actinin. Addition of this sequence module was expected to increase the length of the native α-actinin molecule by at least 15%, yet was fully compatible with α-actinin function as measured by rescued lethality and flight. Thus, unexpectedly, the exact length of α-actinin was not critical to its function in the muscle Z disk. Kluwer Academic Publishers, 2000 Dubreuil Ronald Department of Neurobiology, Pharmacology, and Physiology, University of Chicago, 947 E. 58th Street, 60637, Chicago, IL aut Wang Ping Department of Neurobiology, Pharmacology, and Physiology, University of Chicago, 947 E. 58th Street, 60637, Chicago, IL aut Journal of Muscle Research & Cell Motility Kluwer Academic Publishers 21/7(2000-10-01), 705-713 0142-4319 21:7<705 2000 21 10974 https://doi.org/10.1023/A:1005699608893 text/html Onlinezugriff via DOI 1 research-article jats NATIONALLICENCE 856 40 https://doi.org/10.1023/A:1005699608893 text/html Onlinezugriff via DOI NATIONALLICENCE 700 1- Dubreuil Ronald Department of Neurobiology, Pharmacology, and Physiology, University of Chicago, 947 E. 58th Street, 60637, Chicago, IL aut NATIONALLICENCE 700 1- Wang Ping Department of Neurobiology, Pharmacology, and Physiology, University of Chicago, 947 E. 58th Street, 60637, Chicago, IL aut NATIONALLICENCE 773 0- Journal of Muscle Research & Cell Motility Kluwer Academic Publishers 21/7(2000-10-01), 705-713 0142-4319 21:7<705 2000 21 10974 Metadata rights reserved Springer special CC-BY-NC licence nationallicence BK010053 XK010053 XK010000 NATIONALLICENCE NATIONALLICENCE NL-springer