A single gene encodes the catalytic "A" subunit of the bovine vacuolar H(+)-ATPase.
We have previously demonstrated that the 73-kDa (A) subunit of the bovine coated vesicle (H+)-ATPase possesses a nucleotide binding site required for catalytic activity (Arai, H., Berne, M., Terres, G., Terres, H., Puopolo, K., and Forgac, M. (1987) Biochemistry 26, 6632-6638). Here we report the cDNA sequence of the coding region of the bovine brain A subunit. Comparison of the deduced amino acid sequence with those previously reported for the A subunits of vacuolar ATPases from lower eukaryotes, plants, and archaebacteria reveals significant homology, especially in sequences implicated in nucleotide binding. The message encoding the bovine brain A subunit is relatively large, approximately 4.6 kilobases; Northern blotting of RNA isolated from rat brain and human brain tumor cells reveals a message of similar size. Northern analysis of several bovine tissues indicates that only one message for this subunit is expressed. Southern blot analysis of bovine genomic DNA indicates that the bovine A subunit is encoded by a single gene.
Puopolo
K
K
Department of Physiology, Tufts University School of Medicine, Boston, Massachusetts 02111.
Kumamoto
C
C
Adachi
I
I
Forgac
M
M
D000595:Amino Acid Sequence; D000818:Animals; D001483:Base Sequence; D001665:Binding Sites; D015152:Blotting, Northern; D015139:Blotting, Southern; D001921:Brain / Q000201:enzymology; D002384:Catalysis; D002417:Cattle; D002460:Cell Line; D004247:DNA / Q000235:genetics; D004285:Dogs; D006801:Humans; D008969:Molecular Sequence Data; D016133:Polymerase Chain Reaction; D006180:Proton-Translocating ATPases / Q000235:genetics* / Q000378:metabolism; D012313:RNA / Q000235:genetics; D015183:Restriction Mapping; D016415:Sequence Alignment; D014617:Vacuoles / Q000201:enzymology*
D012313:RNA; D004247:DNA; D006180:Proton-Translocating ATPases