Functional recovery of troponin I in a Drosophila heldup mutant after a second site mutation.
To identify proteins that interact in vivo with muscle components we have used a genetic approach based on the isolation of suppressors of mutant alleles of known muscle components. We have applied this system to the case of troponin I (TnI) in Drosophila and its mutant allele heldup2 (hdp2). This mutation causes an alanine to valine substitution at position 116 after a single nucleotide change in a constitutive exon. Among the isolated suppressors, one of them results from a second site mutation at the TnI gene itself. Muscles endowed with TnI mutated at both sites support nearly normal myofibrillar structure, perform notably well in wing beating and flight tests, and isolated muscle fibers produce active force. We show that the structural and functional recovery in this suppressor does not result from a change in the stoichiometric ratio of TnI isoforms. The second site suppression is due to a leucine to phenylalanine change within a heptameric leucine string motif adjacent to the actin binding domain of TnI. These data evidence a structural and functional role for the heptameric leucine string that is most noticeable, if not specific, in the indirect flight muscle.
Prado
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A
Instituto Cajal, Consejo Superior de Investigaciones Cientificas, Madrid, Spain.
Canal
I
I
Barbas
J A
JA
Molloy
J
J
Ferrús
A
A
D000483:Alleles; D000595:Amino Acid Sequence; D000818:Animals; D001483:Base Sequence; D017931:DNA Primers; D004330:Drosophila / Q000235:genetics / Q000502:physiology*; D005260:Female; D005426:Flight, Animal; D017344:Genes, Insect; D008297:Male; D008854:Microscopy, Electron; D008969:Molecular Sequence Data; D009132:Muscles / Q000502:physiology / Q000648:ultrastructure; D016297:Mutagenesis, Site-Directed; D009210:Myofibrils / Q000502:physiology / Q000648:ultrastructure; D010641:Phenotype; D017354:Point Mutation; D016133:Polymerase Chain Reaction; D013489:Suppression, Genetic; D014336:Troponin / Q000096:biosynthesis* / Q000235:genetics*; D019210:Troponin I
D017931:DNA Primers; D014336:Troponin; D019210:Troponin I