FIGURE 5.

Pol III* with an inactive ATPase cannot support lagging strand DNA synthesis. A, total amount of DNA synthesis was compared between rolling circle reactions containing wild-type Pol III* (DnaX complex (τ₂γ)) and reactions containing mutant Pol III* (DnaX complex (τ₂γ^M)). Rolling circle reactions were conducted in the presence of α-[³²P]dATP and the indicated amount of dGTP. B, α-[³²P]dCTP-, α-[³²P]dGTP-, and α-[³²P]dATP-labeled reaction products from rolling circle reactions were loaded onto an alkaline-agarose gel to assess lagging strand synthesis from wild-type Pol III* (DnaX complex (τ₂γδδ'χψ)) and mutant Pol III* (DnaX complex (τ₂γ^M δδ'χψ)). α-[³²P]dCTP selectively labels the leading strand, α-[³²P]dGTP selectively labels the lagging strand, and α-[³²P]dATP labels both the leading and lagging strands. M, mutant.