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

✍ Scribed by Sarjeet S. Gill; Phillip B. Chu; Peter Smethurst; Patricia V. Pietrantonio; Linda S. Ross

• Book ID: 101291219
• Publisher: John Wiley and Sons
• Year: 1998
• Tongue: English
• Weight: 393 KB
• Volume: 37
• Category: Article
• ISSN: 0739-4462
• DOI: 10.1002/(sici)1520-6327(1998)37:1<80::aid-arch10>3.0.co;2-6

✦ Synopsis

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.