Characterisation of missense mutations in the Act88F gene of Drosophila melanogaster

Douglas R. Drummond, Emma S. Hennessey, John C. Sparrow

Research output: Contribution to journalArticle

99 Citations (Scopus)

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 languageEnglish
Pages (from-to)70-80
Number of pages11
JournalMGG Molecular & General Genetics
Volume226
Issue number1-2
DOIs
Publication statusPublished - Apr 1 1991
Externally publishedYes

Fingerprint

Missense Mutation
Drosophila melanogaster
Actins
Genes
Myofibrils
Heat-Shock Proteins
Diptera
Muscles
Mutagenesis
Polymerization
Detergents
Temperature

All Science Journal Classification (ASJC) codes

  • Genetics

Cite this

Characterisation of missense mutations in the Act88F gene of Drosophila melanogaster. / Drummond, Douglas R.; Hennessey, Emma S.; Sparrow, John C.

In: MGG Molecular & General Genetics, Vol. 226, No. 1-2, 01.04.1991, p. 70-80.

Research output: Contribution to journalArticle

@article{f31c9ee3f8a4490a99dc387397914f6a,
title = "Characterisation of missense mutations in the Act88F gene of Drosophila melanogaster",
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.",
author = "Drummond, {Douglas R.} and Hennessey, {Emma S.} and Sparrow, {John C.}",
year = "1991",
month = "4",
day = "1",
doi = "10.1007/BF00273589",
language = "English",
volume = "226",
pages = "70--80",
journal = "Molecular Genetics and Genomics",
issn = "1617-4615",
publisher = "Springer Verlag",
number = "1-2",

}

TY - JOUR

T1 - Characterisation of missense mutations in the Act88F gene of Drosophila melanogaster

AU - Drummond, Douglas R.

AU - Hennessey, Emma S.

AU - Sparrow, John C.

PY - 1991/4/1

Y1 - 1991/4/1

N2 - 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.

AB - 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.

UR - http://www.scopus.com/inward/record.url?scp=0025794119&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=0025794119&partnerID=8YFLogxK

U2 - 10.1007/BF00273589

DO - 10.1007/BF00273589

M3 - Article

C2 - 1851957

AN - SCOPUS:0025794119

VL - 226

SP - 70

EP - 80

JO - Molecular Genetics and Genomics

JF - Molecular Genetics and Genomics

SN - 1617-4615

IS - 1-2

ER -