Dominant Cold-Sensitive Out-Cold Mutants of Drosophila melanogaster Have Novel Missense Mutations in the Voltage-Gated Sodium Channel Gene paralytic

Here we report the molecular characterization of Out-cold (Ocd) mutants of Drosophila melanogaster, which produce a dominant, X-linked, cold-sensitive paralytic phenotype. From its initial 1.5-Mb cytological location within 13F1-16A2, P-element and SNP mapping reduced the Ocd critical region to <...

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Published inGenetics (Austin) Vol. 180; no. 2; pp. 873 - 884
Main Authors Lindsay, Helen A, Baines, Richard, ffrench-Constant, Richard, Lilley, Kathryn, Jacobs, Howard T, O'Dell, Kevin M.C
Format Journal Article
LanguageEnglish
Published United States Genetics Soc America 01.10.2008
Genetics Society of America
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Abstract Here we report the molecular characterization of Out-cold (Ocd) mutants of Drosophila melanogaster, which produce a dominant, X-linked, cold-sensitive paralytic phenotype. From its initial 1.5-Mb cytological location within 13F1-16A2, P-element and SNP mapping reduced the Ocd critical region to <100 kb and to six candidate genes: hangover, CG9947, CG4420, eIF2a, Rbp2, and paralytic (para). Complementation testing with para null mutations strongly suggests Ocd and para are allelic, as does gene rescue of Ocd semilethality with a wild-type para transgene. Pesticide resistance and electrophysiological phenotypes of Ocd mutants support this conclusion. The para gene encodes a voltage-gated sodium channel. Sequencing the Ocd lines revealed mutations within highly conserved regions of the para coding sequence, in the transmembrane segment S6 of domain III (I1545M and T1551I), and in the linker between domains III and IV (G1571R), the location of the channel inactivation gate. The G1571R mutation is of particular interest as mutations of the orthologous residue (G1306) in the human skeletal muscle sodium channel gene SCN4A are associated with cases of periodic paralysis and myotonia, including the human cold-sensitive disorder paramyotonia congenita. The mechanisms by which sodium channel mutations cause cold sensitivity are not well understood. Therefore, in the absence of suitable vertebrate models, Ocd provides a system in which genetic, molecular, physiological, and behavioral tools can be exploited to determine mechanisms underlying sodium channel periodic paralyses.
AbstractList Here we report the molecular characterization of Out-cold (Ocd) mutants of Drosophila melanogaster, which produce a dominant, X-linked, cold-sensitive paralytic phenotype. From its initial 1.5-Mb cytological location within 13F1-16A2, P-element and SNP mapping reduced the Ocd critical region to <100 kb and to six candidate genes: hangover, CG9947, CG4420, eIF2a, Rbp2, and paralytic (para). Complementation testing with para null mutations strongly suggests Ocd and para are allelic, as does gene rescue of Ocd semilethality with a wild-type para transgene. Pesticide resistance and electrophysiological phenotypes of Ocd mutants support this conclusion. The para gene encodes a voltage-gated sodium channel. Sequencing the Ocd lines revealed mutations within highly conserved regions of the para coding sequence, in the transmembrane segment S6 of domain III (I1545M and T1551I), and in the linker between domains III and IV (G1571R), the location of the channel inactivation gate. The G1571R mutation is of particular interest as mutations of the orthologous residue (G1306) in the human skeletal muscle sodium channel gene SCN4A are associated with cases of periodic paralysis and myotonia, including the human cold-sensitive disorder paramyotonia congenita. The mechanisms by which sodium channel mutations cause cold sensitivity are not well understood. Therefore, in the absence of suitable vertebrate models, Ocd provides a system in which genetic, molecular, physiological, and behavioral tools can be exploited to determine mechanisms underlying sodium channel periodic paralyses.
Initial investigations into the molecular basis of the Ocd cold-sensitive paralysis involved the study of the reaction kinetics of the mitochondrial enzyme complex succinate cytochrome c reductase (SCCR), which reflects the combined enzyme activity of complexes II and III of the respiratory chain (Sondergaard et al. 1975). Fly stocks were maintained in plastic vials or bottles (for large-scale amplification) containing food medium 1% (w/v) bacto-agar, 1.5% (w/v) sucrose, 3% (w/v) glucose, 3.5% (w/v) active dried yeast, 1.\n Interestingly, mutations in the orthologous residue, G1306, in the human skeletal muscle sodium channel SCN4A, have already been found in cases of the human cold-sensitive muscle disorder paramyotonia congenita (G1306V) (McClatchey et al. 1992).
Here we report the molecular characterization of Out-cold ( Ocd ) mutants of Drosophila melanogaster , which produce a dominant, X-linked, cold-sensitive paralytic phenotype. From its initial 1.5-Mb cytological location within 13F1-16A2, P -element and SNP mapping reduced the Ocd critical region to <100 kb and to six candidate genes: hangover , CG9947, CG4420, eIF2a , Rbp2 , and paralytic ( para ). Complementation testing with para null mutations strongly suggests Ocd and para are allelic, as does gene rescue of Ocd semilethality with a wild-type para transgene. Pesticide resistance and electrophysiological phenotypes of Ocd mutants support this conclusion. The para gene encodes a voltage-gated sodium channel. Sequencing the Ocd lines revealed mutations within highly conserved regions of the para coding sequence, in the transmembrane segment S6 of domain III (I1545M and T1551I), and in the linker between domains III and IV (G1571R), the location of the channel inactivation gate. The G1571R mutation is of particular interest as mutations of the orthologous residue (G1306) in the human skeletal muscle sodium channel gene SCN4A are associated with cases of periodic paralysis and myotonia, including the human cold-sensitive disorder paramyotonia congenita . The mechanisms by which sodium channel mutations cause cold sensitivity are not well understood. Therefore, in the absence of suitable vertebrate models, Ocd provides a system in which genetic, molecular, physiological, and behavioral tools can be exploited to determine mechanisms underlying sodium channel periodic paralyses.
Author ffrench-Constant, Richard
O'Dell, Kevin M.C
Lilley, Kathryn
Lindsay, Helen A
Jacobs, Howard T
Baines, Richard
AuthorAffiliation IBLS Division of Molecular Genetics, University of Glasgow, Glasgow G11 6NU, United Kingdom, † Institute of Medical Technology and Tampere University Hospital, FI-33014 University of Tampere, FI-33014 Tampere, Finland, ‡ Faculty of Life Sciences, University of Manchester, Manchester M13 9PT, United Kingdom, § Centre for Ecology and Conservation, Cornwall Campus, University of Exeter, Penryn, Cornwall TR10 9EZ, United Kingdom and Cambridge Centre for Proteomics, Cambridge System Biology Centre, Department of Biochemistry, University of Cambridge, Cambridge CB2 1QR, United Kingdom
AuthorAffiliation_xml – name: IBLS Division of Molecular Genetics, University of Glasgow, Glasgow G11 6NU, United Kingdom, † Institute of Medical Technology and Tampere University Hospital, FI-33014 University of Tampere, FI-33014 Tampere, Finland, ‡ Faculty of Life Sciences, University of Manchester, Manchester M13 9PT, United Kingdom, § Centre for Ecology and Conservation, Cornwall Campus, University of Exeter, Penryn, Cornwall TR10 9EZ, United Kingdom and Cambridge Centre for Proteomics, Cambridge System Biology Centre, Department of Biochemistry, University of Cambridge, Cambridge CB2 1QR, United Kingdom
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Present address: Yorkshire Regional DNA Laboratory, St James's University Hospital, Leeds, UK, LS9 7TF.
Communicating editor: R. Anholt
Corresponding author: IBLS Division of Molecular Genetics, Anderson College Complex, University of Glasgow, 56 Dumbarton Rd., Glasgow G11 6NU, UK. E-mail: k.odell@bio.gla.ac.uk
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SSID ssj0006958
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Snippet Here we report the molecular characterization of Out-cold (Ocd) mutants of Drosophila melanogaster, which produce a dominant, X-linked, cold-sensitive...
Initial investigations into the molecular basis of the Ocd cold-sensitive paralysis involved the study of the reaction kinetics of the mitochondrial enzyme...
Here we report the molecular characterization of Out-cold ( Ocd ) mutants of Drosophila melanogaster , which produce a dominant, X-linked, cold-sensitive...
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proquest
crossref
pubmed
highwire
fao
SourceType Open Access Repository
Aggregation Database
Index Database
Publisher
StartPage 873
SubjectTerms Alleles
Amino Acid Sequence
Animals
cold
dominant genes
Drosophila melanogaster
Drosophila melanogaster - genetics
Drosophila melanogaster - physiology
Drosophila Proteins - genetics
Drosophila Proteins - metabolism
Genes
Genes, Insect
Genetic disorders
Genetics
Genotype & phenotype
Investigations
Males
Mass spectrometry
missense mutation
Mitochondrial Proteins - genetics
Mitochondrial Proteins - metabolism
Models, Biological
Molecular Sequence Data
mutants
Mutation
Mutation, Missense
Ocd gene
Out-cold mutants
para gene
Paralyses, Familial Periodic - genetics
Paralyses, Familial Periodic - metabolism
paralysis
Phenotype
Proteins
Reaction kinetics
sodium channels
Sodium Channels - genetics
Sodium Channels - metabolism
Studies
Thermosensing - genetics
Title Dominant Cold-Sensitive Out-Cold Mutants of Drosophila melanogaster Have Novel Missense Mutations in the Voltage-Gated Sodium Channel Gene paralytic
URI http://www.genetics.org/cgi/content/abstract/180/2/873
https://www.ncbi.nlm.nih.gov/pubmed/18723887
https://www.proquest.com/docview/214133466
https://search.proquest.com/docview/69671294
https://pubmed.ncbi.nlm.nih.gov/PMC2567387
Volume 180
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