Isolation of 88F Actin Mutants of Drosophila melanogasterand Possible Alterations in the Mutant Actin Structures

• Published in: Journal of molecular biology 1996, Vol.260 (4), p.492-505
• Main Authors: An, HyeSuck, Mogami, Kaname
• Format: Article
• Language: English
• Subjects: Drosophila; actin; mutant; PCR; protein folding
• ISSN: 0022-2836; 1089-8638
• DOI: 10.1006/jmbi.1996.0417

The 88F actin (act88F) gene of Drosophila melanogasterencodes an actin isoform that is expressed exclusively in the indirect flight muscle. In order to isolate a large number of act88F mutants, an efficient screening method was used to obtain dominant flightless mutants. Genetic analyses revealed that 25 mutations were located near or at the act88F locus. From each mutant strain, the DNA fragments including the coding region of the act88F gene were asymmetrically amplified by the polymerase chain reaction method, and the amplified fragments were directly sequenced. Eighteen of them were found to have point mutations within their coding regions. Of these, 13 were novel alleles of this gene. We have characterised these mutations in detail. First, their flight abilities were tested after introducing two normal alleles of this gene. Second, two-dimensional gel electrophoresis was used to examine actin isoforms and whole thorax proteins. Third, morphological anomalies of indirect flight muscle fibres and myofibrils were examined with an optical microscope. On the basis of these phenotypes and the known atomic structure of actin, possible alterations in the structure of actin brought about by these mutations are discussed.