Supplemental Material

This article contains the following supporting material:

• Table 1
• Table 2
• Figure 1 - GFP tagged rad4^+TopBP1 or rad4-c11^TopBP1 has identical phenotype as the untagged cells.
  (A) Expression of rad4^+TopBP1 or rad4^+TopBP1:GFP was able to restore temperature sensitivity of rad4-116^TopBP1. Strain leu1-32 was independently transformed with plasmid, pREP41 and used as empty vector control. rad4-116^TopBP1 strain was independently transformed with plasmids, pREP41 (empty vector control), prad4^+TopBP1, and prad4^+TopBP1:GFP. Transformants (1x10⁷) were serial diluted (1:10), spotted on EMM –LEU plates and incubated at 30°C and 36°C for 5 days. (B) GFP-tag of rad4^+TopBP1 and rad4-c11^+TopBP1 does not alter the sensitivities of cells to genotoxic agents. Cells (1x10⁷) were cultured to log phase, 10-fold serial diluted, and spotted onto YES plates or YES plates with 5 mM HU and 10 μM CPT and incubated at 30°C for 3 days.

• Figure 2 - Detection of Crb3 protein sequence in Rad4^TopBP1:GFP immunoprecipitates by the tandem mass spectrometric (MS/MS) analysis.
  (A) Rad4^TopBP1:GFP immunoprecipitates, after CPT treatment as described in Material & Methods revealed a peptide fragment having sequence, LYTASEDNTIR, matching the Crb3 protein sequence. (B) No peptide fragment corresponding to the Crb3 protein sequence was detected in Rad4-c11^TopBP1:GFP immunoprecipitates, after CPT treatment as described in Materials & Methods.