FIGURE 5.

A, effects of dATP and ATP on RNA and DNA synthesis. Mixtures (20 μl), as indicated under “Experimental Procedures,” contained 100 μm of each of the four dNTPs or rNTPs with [α-³²P]dATP (1344 cpm/pmol) or [α-³²P]ATP (1357 cpm/pmol), 83.4 fmol of M13 DNA, 5.6 pmol of the T. kodakaraensis primase complex, and varying levels of unlabeled dATP or ATP (1000, 300, 100, 30, and 10 μm), where indicated. After 20 min at 60 °C, products were subjected to urea-PAGE analysis, and the gels were exposed for 1 h at −70 °C. M, molecular mass markers; inc., incorporation. B, influence of ATP, dATP, GTP, and dGTP on polynucleotide synthesis in reactions containing oligo(dT)₃₀ as template. Reactions were as described in the legend for Fig. 3 with 10 pmol of oligo(dT)₃₀. Mixtures contained 100 μm [α-³²P]dATP (6815 cpm/pmol) or 100 μm [α-³²P]ATP (6024 cpm/pmol) and varying levels of ATP (1000, 100, and 10 μm), dATP (1000, 100, and 10 μm), GTP (1000 μm), or dGTP (1000 μm) where indicated. Following incubation, 5 μl of each reaction were subjected to urea-PAGE analysis and autoradiography. Reaction mixtures in lanes 15 and 16 contained 100 μm [α-³²P]rGTP (10⁴ cpm/pmol) or 100 μm [³²P]dGTP (10⁴ cpm/pmol), respectively. comp., complex.