Figure 5
Analysis of the nucleotide-linked protein in the presence of different combinations of deoxynucleotides and dideoxynucleotides. Micrococcal nuclease (MN)-treated pFOXC3-containing mitochondrial ribonucleoproteins (mtRNPs) were incubated with [α-32P]dATP or [α-32P]deoxyguanosine triphosphate (dGTP), and with unlabeled deoxyribonucleotide triphosphates (dNTPs) or dideoxynucleotide triphosphates (ddNTPs), as indicated. Reactions were terminated with Laemmli buffer and reaction products were separated via 4% to 20% SDS-PAGE. Prestained protein size markers are indicated on the left in kDa. (a) Reactions having [α-32P]dATP or [α-32P]dGTP with one unlabeled deoxynucleotide. (b) Reactions having [α-32P]dATP or [α-32P]dGTP, with a full complement of deoxynucleotides or dideoxynucleotides. (c) Reactions having [α-32P]dGTP and a single dideoxynucleotide with a complement of deoxynucleotides, as indicated. (d) Reactions having [α-32P]dATP and a full complement of deoxynucleotides incubated with or without phosphonoformate (PFA) at the indicated concentrations. Prestained protein size markers are indicated on the left in kDa and bands showing a slight size difference are indicated by an arrow.