Monoubiquitination of PCNA does not change the pattern of TLS by DNA polymerase λ.

<p>Experiments were performed with a damaged template primed with the 74 mer primer, as depicted on the top of the Fig 5A and in <a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0093908#pone-0093908-g001" target="_blank">Fig 1</a>. The proteins are indicated at the top of the figure. Experiments were carried out as described in Materials and Methods. <b>A</b>: lane 1, no proteins present. Lane 2, reaction incubated for 35 minutes with 0.025 pmol of pol ε. Lane 3, reaction incubated for 35 minutes with 0.025 pmol of pol ε, 0.25 pmol of RPA and 160 ngs of RFC. Lane 4, reaction incubated for 30 minutes with 0.025 pmol of pol ε, 0.25 pmol of RPA and 160 ngs of RFC; then 0.25 pmol of pol λ was added and the incubation continued for 5 minutes. Lanes 5 to 7; as in lane 4 but with 0.3, 0.6 and 1.2 pmol of PCNA respectively. Lanes 8 to 10; as in lane 4 but with 0.3, 0.6 and 1.2 pmol of monoubiquitinated PCNA respectively. The position of the primers, the AP site and the nucleotides past the AP site are indicated. <b>B</b>: Quantification of the percentage of TLS, calculated as described in Materials and Methods. Mean +/− S.D. values for three independent experiments are indicated.