Drosophila, Muscle Regulation Characterized by Electron Microscopy and Three-Dimensional Reconstruction of Thin Filament Mutants

Anthony Cammarato, Victoria Hatch, Judith Saide, Roger Craig, John C. Sparrow, Larry S. Tobacman, William Lehman

Research output: Contribution to journalArticle

Abstract

Wild-type and mutant thin filaments were isolated directly from "myosinless" Drosophila indirect flight muscles to study the structural basis of muscle regulation genetically. Negatively stained filaments showed tropomyosin with periodically arranged troponin complexes in electron micrographs. Three-dimensional helical reconstruction of wild-type filaments indicated that the positions of tropomyosin on actin in the presence and absence of Ca2+ were indistinguishable from those in vertebrate striated muscle and consistent with a steric mechanism of regulation by troponin-tropomyosin in Drosophila muscles. Thus, the Drosophila model can be used to study steric regulation. Thin filaments from the Drosophila mutant heldup2, which possesses a single amino acid conversion in troponin 1, were similarly analyzed to assess the Drosophila model genetically. The positions of tropomyosin in the mutant filaments, in both the Ca 2+-free and the Ca2+-induced states, were the same, and identical to that of wild-type filaments in the presence of Ca2+. Thus, cross-bridge cycling would be expected to proceed uninhibited in these fibers, even in relaxing conditions, and this would account for the dramatic hypercontraction characteristic of these mutant muscles. The interaction of mutant troponin I with Drosophila troponin C is discussed, along with functional differences between troponin C from Drosophila and vertebrates.

Original languageEnglish (US)
Pages (from-to)1618-1624
Number of pages7
JournalBiophysical journal
Volume86
Issue number3
DOIs
StatePublished - Mar 2004
Externally publishedYes

ASJC Scopus subject areas

  • Biophysics

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