Myosin light chain-2 mutation affects flight, wing beat frequency, and indirect flight muscle contraction kinetics in Drosophila.
We have used a combination of classical genetic, molecular genetic, histological, biochemical, and biophysical techniques to identify and characterize a null mutation of the myosin light chain-2 (MLC-2) locus of Drosophila melanogaster. Mlc2E38 is a null mutation of the MLC-2 gene resulting from a nonsense mutation at the tenth codon position. Mlc2E38 confers dominant flightless behavior that is associated with reduced wing beat frequency. Mlc2E38 heterozygotes exhibit a 50% reduction of MLC-2 mRNA concentration in adult thoracic musculature, which results in a commensurate reduction of MLC-2 protein in the indirect flight muscles. Indirect flight muscle myofibrils from Mlc2E38 heterozygotes are aberrant, exhibiting myofilaments in disarray at the periphery. Calcium-activated Triton X-100-treated single fiber segments exhibit slower contraction kinetics than wild type. Introduction of a transformed copy of the wild type MLC-2 gene rescues the dominant flightless behavior of Mlc2E38 heterozygotes. Wing beat frequency and single fiber contraction kinetics of a representative rescued line are not significantly different from those of wild type. Together, these results indicate that wild type MLC-2 stoichiometry is required for normal indirect flight muscle assembly and function. Furthermore, these results suggest that the reduced wing beat frequency and possibly the flightless behavior conferred by Mlc2E38 is due in part to slower contraction kinetics of sarcomeric regions devoid or partly deficient in MLC-2.
Warmke
J
J
Department of Molecular Genetics, Ohio State University, Columbus 43210.
Yamakawa
M
M
Molloy
J
J
Falkenthal
S
S
Maughan
D
D
D000494:Allosteric Regulation; D000595:Amino Acid Sequence; D000818:Animals; D001483:Base Sequence; D000252:Calcium-Transporting ATPases / Q000378:metabolism; D002874:Chromosome Mapping; D004171:Diploidy; D004331:Drosophila melanogaster / Q000502:physiology*; D005426:Flight, Animal / Q000502:physiology*; D006579:Heterozygote; D007700:Kinetics; D008969:Molecular Sequence Data; D009119:Muscle Contraction / Q000502:physiology*; D009132:Muscles / Q000002:abnormalities / Q000502:physiology* / Q000648:ultrastructure; D009154:Mutation; D009210:Myofibrils / Q000648:ultrastructure; D009218:Myosins / Q000096:biosynthesis / Q000235:genetics*; D014921:Wings, Animal / Q000502:physiology
D009218:Myosins; D000252:Calcium-Transporting ATPases