Isolation of the V-ATPase A and c subunit cDNAs from mosquito midgut and Malpighian tubules.

Autor(es): Gill S S; Chu P B; Smethurst P; Pietrantonio P V; Ross L S

Resumo: 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.

Palavras-Chave: V-ATPase; A subunit c subunit Aedes aegypti; cDNA clones midgut; Malpighian tubules

Imprenta: Archives of Insect Biochemistry and Physiology, v. 37, n. 1, p. 80-90, 1998

Identificador do objeto digital: 10.1002/(SICI)1520-6327(1998)37:1<80::AID-ARCH10>3.0.CO;2-6

Descritores: Aedes aegypti - DNA ; Aedes aegypti - Genome ; Aedes aegypti - Molecular Structure ; Aedes aegypti - Pathogenesis ; Aedes aegypti - Proteins

Data de publicação: 1998