Isolation of the V-ATPase A and c subunit cDNAs from mosquito midgut and Malpighian tubules.
Using conserved amino acid sequences for the design of oligonucleotide primers, we isolated cDNA clones for two subunits of the V-ATPase from the midgut and Malpighian tubules of Aedes aegypti larvae. The 3.1 kb cDNA of the A subunit of the peripheral catalytic V1 sector codes for a protein of 68.6 kDa. The protein contains conserved motifs, including an ATP/GTP binding site, found in all other A subunits. Southern analysis using the A subunit as a probe suggests the presence of only a single copy of gene in the Aedes aegypti. The 0.85 kb cDNA of the c subunit of the membrane H+ conducting V0 sector codes for a protein of kDa. This protein has four transmembrane domains and contains a conserved glutamic acid that serves as the binding site for dicyclohexylcarbodiimide. Southern analysis using the c subunit as a probe suggests the presence of more than one related gene in the genome of Aedes aegypti. Pileup analysis of various A and c subunits shows that these subunits fall into distinct clusters, including one in which all arthropod proteins are clustered.
Gill
S S
SS
Department of Entomology, University of California, Riverside 92521, USA. Gill@ucrac1.ucr.edu
Chu
P B
PB
Smethurst
P
P
Pietrantonio
P V
PV
Ross
L S
LS
D000330:Aedes / Q000196:embryology / Q000201:enzymology / Q000235:genetics*; D000595:Amino Acid Sequence; D000818:Animals; D001483:Base Sequence; D018076:DNA, Complementary; D007814:Larva / Q000201:enzymology; D008317:Malpighian Tubules / Q000201:enzymology*; D008969:Molecular Sequence Data; D006180:Proton-Translocating ATPases / Q000235:genetics*; D013045:Species Specificity; D025262:Vacuolar Proton-Translocating ATPases
D018076:DNA, Complementary; D025262:Vacuolar Proton-Translocating ATPases; D006180:Proton-Translocating ATPases