Abstract
Bacterial operons for F(1)F(o)-ATP synthase typically include an uncI gene that encodes a function-unknown small hydrophobic protein. When we expressed a hybrid F(1)F(o) (F(1) from thermophilic Bacillus PS3 and Na(+)-translocating F(o) from Propionigenium modestum) in Escherchia coli cells, we found that uncI derived from P. modestum was indispensable to produce active enzyme; without uncI, c-subunits in F(1)F(o) existed as monomers but not as functional c(11)-ring. When uncI was expressed from another plasmid at the same time, active F(1)F(o) with c(11)-ring was produced. A plasmid containing only uncI and c-subunit gene produced c(11)-ring, but a plasmid containing only c-subunit gene did not. Direct interaction of UncI protein with c-subunits was suggested from copurification of His-tagged UncI protein and c-subunits, both in the state of c(11)-ring and c-monomers. Na(+) induced dissociation of His-tagged UncI protein from c(11)-ring but not from c-monomers. These results show that UncI is a chaperone-like protein that assists c(11)-ring assembly from c-monomers in the membrane.