A Network of HMG-box Transcription Factors Regulates Sexual Cycle in the Fungus Podospora anserina
Figure 8
Electrophoretic mobility shift assays with PaHMG5.
(A) Interaction of His tagged PaHMG5 (HMG5His) with probes corresponding to FMR1, FMR1 scrambled sequence (FMR1Sc), FPR1, PaHMG8 and PaHMG5 oligonucleotides. The probes are indicated below each panel. The interaction of HMG5His with probe was analyzed without competitor and in the presence of increasing amounts (given as fold molar excess below the triangles) of competitor (indicated above the triangles). (B) Interaction of the HMG5His protein with the MFM probe and reciprocal competition between FMR1 and MFM oligonucleotides. The probes are indicated below each panel. The competitors are indicated above the triangles. A control of the interaction of HMG5His with the FMR1 probe was performed as in A and included in the assay. HMG5His has a greater affinity for FMR1 than for MFM sequence, as indicated by the efficient exclusion of MFM probe by FMR1 competitor and the very inefficient exclusion of FMR1 probe by MFM competitor. (C) Interaction of the HMG5His protein with the MFP probe and reciprocal competition between FMR1 and MFP oligonucleotides. Legend as in B. HMG5His has a greater affinity for FMR1 than for MFP sequence, as indicated by the efficient exclusion of MFP probe by FMR1 competitor and the inefficient exclusion of FMR1 probe by MFP competitor. (D) Interaction of the HMG5His protein with the PRE1 probe and reciprocal competition between FMR1 and PRE1 oligonucleotides. Legend as in B. HMG5His has a greater affinity for FMR1 than for PRE1 oligonucleotides, as indicated by the efficient exclusion of PRE1 probe by FMR1 competitor and the inefficient exclusion of FMR1 probe by PRE1 competitor.