Figure 8.

Rad51-NTD is a tunable ‘nanny’ peptide. The wild-type SK1 haploid strain was transformed with an indicated yeast expression vector, i.e. P_RAD51-LacZ-NLS-V5 (mock), P_RAD51-NTD^WT-LacZ-NLS-V5, P_RAD51-NTD^3A-LacZ-NLS-V5 or P_RAD51-NTD^3D-LacZ-NLS-V5. (A, B) Immunoblotting. Total cell lysates were prepared from mitotic cells undergoing exponential growth for visualization by immunoblotting with the corresponding antisera and quantitative yeast LacZ assays in (C). (A) NTD-LacZ-NLS-V5 fusion proteins (∼128 kDa) are indicated by white arrowheads. Endogenous Rad51 (∼43 kDa) is indicated by black arrows. Numbers below the anti-Rad51 immunoblot indicate fold-changes of NTD-LacZ-NLS-V5 fusion protein levels normalized to endogenous Rad51 protein levels in the same blot. (B) Immunoblotting using anti-V5 antiserum. Presumptively degraded products are indicated by black arrowheads. Hsp104 was used as a loading control. (C) Quantitative yeast LacZ assays were performed as described previously (41). Error bars indicate standard deviation between experiments. (n = 3). Asterisks indicate significant differences when compared to NTD^WT, with P values calculated using a two-tailed t-test (*P value < 0.05 and ***P value < 0.001).