Deletion analysis of the subunit genes of V-type Na+-ATPase from Enterococcus hirae.
The V1Vo-ATPase from Enterococcus hirae catalyzes ATP hydrolysis coupled with sodium translocation. Mutants with deletions of each of 10 subunits (NtpA, B, C, D, E, F, G, H, I, and K) were constructed by insertion of a chloramphenicol acetyltransferase gene into the corresponding subunit gene in the genome. Measurements of cell growth rates, 22Na+ efflux activities, and ATP hydrolysis activities of the membranes of the deletion mutants indicated that V-ATPase requires nine of the subunits, the exception being the NtpH subunit. The results of Western blotting and V1-ATPase dissociation analysis suggested that the A, B, C, D, E, F, and G subunits constitute the V1 moiety, whereas the V0 moiety comprises the I and K subunits.
Hosaka
Toshiaki
T
Genomic Sciences Center, RIKEN, 1-7-22 Suehiro-cho, Tsurumi-ku, Yokohama, Kanagawa 230-0045.
Takase
Kazuma
K
Murata
Takeshi
T
Kakinuma
Yoshimi
Y
Yamato
Ichiro
I
D000251:Adenosine Triphosphatases / Q000235:genetics* / Q000378:metabolism; D000255:Adenosine Triphosphate / Q000378:metabolism; D001426:Bacterial Proteins / Q000235:genetics / Q000378:metabolism; D015153:Blotting, Western; D027682:Cation Transport Proteins / Q000235:genetics* / Q000378:metabolism; D002462:Cell Membrane / Q000378:metabolism; D004591:Electrophoresis, Polyacrylamide Gel; D016983:Enterococcus / Q000201:enzymology* / Q000235:genetics / Q000254:growth & development; D017353:Gene Deletion; D005816:Genetic Complementation Test; D006863:Hydrogen-Ion Concentration; D007150:Immunohistochemistry; D017136:Ion Transport / Q000235:genetics; D008957:Models, Genetic; D016254:Mutagenesis, Insertional / Q000235:genetics / Q000379:methods*; D009154:Mutation / Q000235:genetics; D009876:Operon / Q000235:genetics; D021122:Protein Subunits / Q000235:genetics / Q000378:metabolism; D012964:Sodium / Q000378:metabolism; D025262:Vacuolar Proton-Translocating ATPases / Q000235:genetics* / Q000378:metabolism
D001426:Bacterial Proteins; D027682:Cation Transport Proteins; D021122:Protein Subunits; D000255:Adenosine Triphosphate; D012964:Sodium; D000251:Adenosine Triphosphatases; D025262:Vacuolar Proton-Translocating ATPases; C021838:sodium-translocating ATPase