Flightin is essential for thick filament assembly and sarcomere stability in Drosophila flight muscles.
Flightin is a multiply phosphorylated, 20-kD myofibrillar protein found in Drosophila indirect flight muscles (IFM). Previous work suggests that flightin plays an essential, as yet undefined, role in normal sarcomere structure and contractile activity. Here we show that flightin is associated with thick filaments where it is likely to interact with the myosin rod. We have created a null mutation for flightin, fln(0), that results in loss of flight ability but has no effect on fecundity or viability. Electron microscopy comparing pupa and adult fln(0) IFM shows that sarcomeres, and thick and thin filaments in pupal IFM, are 25-30% longer than in wild type. fln(0) fibers are abnormally wavy, but sarcomere and myotendon structure in pupa are otherwise normal. Within the first 5 h of adult life and beginning of contractile activity, IFM fibers become disrupted as thick filaments and sarcomeres are variably shortened, and myofibrils are ruptured at the myotendon junction. Unusual empty pockets and granular material interrupt the filament lattice of adult fln(0) sarcomeres. Site-specific cleavage of myosin heavy chain occurs during this period. That myosin is cleaved in the absence of flightin is consistent with the immunolocalization of flightin on the thick filament and biochemical and genetic evidence suggesting it is associated with the myosin rod. Our results indicate that flightin is required for the establishment of normal thick filament length during late pupal development and thick filament stability in adult after initiation of contractile activity.
Reedy
M C
MC
Department of Cell Biology, Duke University Medical Center, Durham, North Carolina 27710, USA.
Bullard
B
B
Vigoreaux
J O
JO
D000818:Animals; D001483:Base Sequence; D003001:Cloning, Molecular; D029721:Drosophila Proteins; D004331:Drosophila melanogaster / Q000166:cytology / Q000235:genetics / Q000378:metabolism* / Q000648:ultrastructure; D005298:Fertility; D064448:Filamins; D005426:Flight, Animal; D017353:Gene Deletion; D006579:Heterozygote; D007106:Immune Sera / Q000276:immunology; D008855:Microscopy, Electron, Scanning; D008954:Models, Biological; D009124:Muscle Proteins / Q000235:genetics / Q000276:immunology / Q000378:metabolism*; D018482:Muscle, Skeletal / Q000737:chemistry / Q000166:cytology / Q000378:metabolism* / Q000648:ultrastructure*; D009218:Myosins / Q000378:metabolism; D010641:Phenotype; D011679:Pupa / Q000166:cytology; D012518:Sarcomeres / Q000378:metabolism* / Q000648:ultrastructure*; D012995:Solubility
D029721:Drosophila Proteins; D064448:Filamins; D007106:Immune Sera; D009124:Muscle Proteins; C081072:fln protein, Drosophila; D009218:Myosins