Interactions between MAGE and EID Family Proteins Table 1. Interaction of S.pombe Nse3 mutants with Nse1 and Nse4. Mutation Nse1 Nse4 Nse4 Phenotype WT L93A V94A R95A V98A R99A Y100A I102A Q105A S107A H108A N109A T112A R113A K114A K119A F121A E123A T125A R127A F130A Q131A V133A F134A E135A E136A A137K R139A Q140A L141A S144A F145A G146A F147A L149A V152A S155A H180A Y182A V184A L185A T188A L199A F212A + + 2 + 2 2 + 2 + + + + + + + + 2 + + + +/2 + +/2 2 + + 2 +/2 + 2 + + 2 2 2 + + + + + 2 + + 2 + + + + + + + +/2 + + + + + + + + +/2 + + + + + + + + + + + + +/2 + + +/2 + + + + + + + + + + 2 Phenotype WT WT WT WT WT WT WT Mutation M214A T215A V216A I217A A218G F219A I220A V222A S223A V227A H229A L232A F235A L236A E238A L239A L240A P247A L248A H249A I252A S255A S257A L259A V260A R261A Q262A Y264A L265A R267A F276A Y278A Y279A E287A L293A F296A V297A F300A F301A SUMMARY Nse1 + + + 2 + + 2 + + + + 2 + + + + + + + + + + + + + + 2 2 2 + 2 2 + + 2 + + + + Structural Nse4-specific Nse1-specific No disruption Total 2 +/2 2 2 2 2 2 2 2 + + 2 2 2 + 2 2 2 2 2 2 2 + 2 + + 2 2 2 + + 2 + + 2 2 2 2 + WT WT WT WT WT WT WT WT WT WT WT WT WT 13 20 12 37 82 Combinations of the indicated mutant Nse3 proteins with Nse1 or Nse4 were analysed in the Y2H system. + and 2 signify whether or not an interaction was detected. Some of the mutations were introduced into the S. pombe genome and sensitivity to MMS and HU was analysed. WT indicates no sensitivity. doi:10.1371/journal.pone.0017270.t001 the cells with NSE4b and the series of mutants of MAGEG1 that reduced or abolished its interaction with NSE4b (see Figure 5). As seen in Figure 6A, lanes 9–11 and lanes 17–19, transcriptional activation by the mutant MAGEG1 proteins and NSE4b was much less than with the corresponding concentration of wild-type MAGEG1 (Lanes 8 and 16). PLoS ONE | www.plosone.org 5 February 2011 | Volume 6 | Issue 2 | e17270