Figure 2.

Modulation of PrimPol primase activity by RPA. The inhibitory effect of RPA on PrimPol primase activity was evaluated using two preferred templates differing in their size: (a) 29-mer GTCC or (b) 60-mer GTCC. The primase reaction contained 16 nM [γ-³²P]ATP, dGTP (1, 10, 100 µM), 5 mM MgCl₂ and 1 mM MnCl₂ as metal cofactors, 1 µM of the indicated template, PrimPol (200 nM), and RPA (12, 50, 200 nM) when indicated. (c) RPA stimulates PrimPol primase activity on a heterogeneous 65-mer oligonucleotide. The reaction contained t16 nM [α-³²P]dGTP, 100 µM of dATP, dCTP and dTTP as indicated, 5 mM MgCl₂ and 1 mM MnCl₂ as metal cofactors, PrimPol (200 nM), and RPA (200 nM) when indicated. The schemes depict the template oligonucleotide, PrimPol (grey square), and the synthesized primer product.