Abstract
We have created missense mutations in the indirect flight muscle (IFM)-specific Act88F actin gene of Drosophila melanogaster by random in vitro mutagenesis. Following P element-mediated transformation into wild-type flies and subsequent transfer of the inserts into Act88F null strains, the effects of the actin mutants on the structure and function of the IFMs were examined. All of the mutants were antimorphic for flight ability. E316K and G368E formed muscle with only relatively small defects in structure whilst the others produced IFMs with large amounts of disruption. E334K formed filaments but lacked Z discs. V339I formed no muscle structure in null flies and did not accumulate actin. E364K and G366D both had relatively stable actin but did not form myofibrils. Using an in vitro polymerisation assay we found no significant effects on the ability of the mutant actins to polymerise. E364K and G366D also caused a strong induction of heat shock protein (hsp) synthesis at normal temperatures and accumulated large amounts of hsp22 which, together with the mutant actin, was resistant to detergent extraction. Both E316K and E334K caused a weak induction of hsp synthesis. We discuss how the stability, structure and function of the different mutant actins affects myofibril assembly and function, and the induction of hsps.
| Original language | English |
|---|---|
| Pages (from-to) | 70-80 |
| Number of pages | 11 |
| Journal | MOLECULAR AND GENERAL GENETICS |
| Volume | 226 |
| Issue number | 1-2 |
| Publication status | Published - Apr 1991 |
Keywords
- ACTIN
- DROSOPHILA
- HEAT SHOCK
- MUSCLE
- HEAT-SHOCK PROTEINS
- SITE-SPECIFIC MUTAGENESIS
- INDIRECT FLIGHT MUSCLES
- MUTANT ACTIN GENES
- MYOFIBRILLAR PROTEINS
- ABNORMAL PROTEINS
- INDUCE
- POLYMERIZATION
- ACTIVATION
- EXPRESSION