Substitution of flight muscle-specific actin by human (beta)-cytoplasmic actin in the indirect flight muscle of Drosophila.
The human (beta)-cytoplasmic actin differs by only 15 amino acids from Act88F actin which is the only actin expressed in the indirect flight muscle (IFM) of Drosophila melanogaster. To test the structural and functional significance of this difference, we ectopically expressed (beta)-cytoplasmic actin in the IFM of Drosophila that lack endogenous Act88F. When expression of the heterologous actin was regulated by approximately 1.5 kb of the 5' promoter region of the Act88F gene, little (beta)-cytoplasmic actin accumulated in the IFM of the flightless transformants. Including Act88F-specific 5' and 3' untranslated regions (UTRs) yielded transformants that expressed wild-type amounts of (beta)-cytoplasmic actin. Despite the assembly of (beta)-cytoplasmic actin containing thin filaments to which endogenous myosin crossbridges attached, sarcomere organization was deficient, leaving the transformants flightless. Rather than affecting primarily actin-myosin interactions, our findings suggest that the (beta)-cytoplasmic actin isoform is not competent to interact with other actin-binding proteins in the IFM that are involved in the organization of functional myofibrils.
Brault
V
V
M.E. Müller Institute, Biozentrum, University of Basel, CH-4056 Basel, Switzerland.
Reedy
M C
MC
Sauder
U
U
Kammerer
R A
RA
Aebi
U
U
Schoenenberger
C
C
D000199:Actins / Q000235:genetics*; D000818:Animals; D003593:Cytoplasm / Q000737:chemistry / Q000235:genetics*; D004331:Drosophila melanogaster / Q000502:physiology*; D005426:Flight, Animal / Q000502:physiology*; D005786:Gene Expression Regulation / Q000235:genetics; D006801:Humans; D008854:Microscopy, Electron; D018482:Muscle, Skeletal / Q000737:chemistry* / Q000502:physiology / Q000648:ultrastructure; D016296:Mutagenesis / Q000235:genetics; D011401:Promoter Regions, Genetic / Q000235:genetics; D020033:Protein Isoforms / Q000235:genetics; D019076:Transgenes / Q000235:genetics
D000199:Actins; D020033:Protein Isoforms