Fig. 9
The DNA-binding domain (DBD) of PoxCxrA. a Schematic diagram of the PoxCxrA protein containing a Zn2Cys6 domain, and DBD truncation. b The minimal DBD11–58 determined via in vitro EMSA. c Interaction between PoxCxrA11–58 and PBM_POX05587. d Interaction between PoxCxrA17–58 and PBM_POX05587. Each EMSA experiment comprised 2 µg of the truncated PoxCxrA-binding domain and 50 ng of PBM_POX05587 labelled with 6-carboxyfluorescein. The promoter of β-tubulin gene POX05989 and PBM2 were used as control probes. The fusion protein Trx–His–S purified from E. coli cells harbouring the empty plasmid pET32a (+), and BSA alone, were used as protein controls. ‘−’ and ‘+’ represent EMSA without and with DBD of PoxCxrA11–58. ‘*’indicated the mutated amino acids into alanine in the mutated DBD of PoxCxrA11–58. e Alignment of DBD17–58 with known Zn2Cys6-containing transcription factors; PoxCxrA and PoxClrB are from P. oxalicum strain HP7-1 (accession numbers KY368172 and KU597415); Pho7, XlnR, Clr1 and Gal4 are from Schizosaccharomyces pombe, P. oxalicum strain 114-2 (EPS32714), N. crassa strain OR74A (XP_011394265) and Saccharomyces cerevisiae S288C (NP_015076), respectively. f Interaction between the mutated PoxCxrA17–58 and PBM_POX05587