Molecular genetic analysis of V-ATPase function in Drosophila melanogaster.
V-ATPases are phylogenetically widespread, highly conserved, multisubunit proton pumps. Originally characterised in endomembranes, they have been found to energise transport across plasma membranes in a range of animal cells and particularly in certain epithelia. While yeast is the model of choice for the rapid generation and identification of V-ATPase mutants, it does not allow their analysis in a plasma membrane context. For such purposes, Drosophila melanogaster is a uniquely suitable model. Accordingly, we have cloned and characterised genes encoding several V-ATPase subunits in D. melanogaster and, using P-element technology, we have succeeded in generating multiple new alleles. Reporter gene constructs reveal ubiquitous expression, but at particularly high levels in those epithelial thought to be energised by V-ATPases, and several of the alleles have lethal recessive phenotypes characterised by epithelial dysfunction. These results, while providing the first gene knockouts of V-ATPases in animals, also illustrate the general utility of D. melanogaster as a model for the genetic analysis of ion transport and its control in epithelia.
Dow
J A
JA
Division of Molecular Genetics, University of Glasgow, UK. j.a.t.dow@bio.gla.ac.uk
Davies
S A
SA
Guo
Y
Y
Graham
S
S
Finbow
M E
ME
Kaiser
K
K
D000483:Alleles; D000818:Animals; D002462:Cell Membrane / Q000201:enzymology; D003001:Cloning, Molecular; D029721:Drosophila Proteins; D004331:Drosophila melanogaster / Q000235:genetics* / Q000378:metabolism; D004790:Enzyme Induction; D018390:Gene Targeting; D017344:Genes, Insect; D005804:Genes, Lethal; D017930:Genes, Reporter; D019476:Insect Proteins / Q000378:metabolism; D007425:Intracellular Membranes / Q000201:enzymology; D008317:Malpighian Tubules / Q000201:enzymology; D016254:Mutagenesis, Insertional; D009928:Organ Specificity; D063150:Polycomb Repressive Complex 1; D017494:Proton Pumps / Q000235:genetics* / Q000502:physiology; D018892:Proton-Motive Force; D006180:Proton-Translocating ATPases / Q000235:genetics* / Q000502:physiology; D025262:Vacuolar Proton-Translocating ATPases
D029721:Drosophila Proteins; D019476:Insect Proteins; C066770:Pc protein, Drosophila; D017494:Proton Pumps; D063150:Polycomb Repressive Complex 1; D025262:Vacuolar Proton-Translocating ATPases; D006180:Proton-Translocating ATPases