Transcriptional Activation of the Aspergillus nidulans gpdA Promoter by Osmotic Signals

ABSTRACT A differentially expressed gpdA cDNA clone was isolated from NaCl-adapted Aspergillus nidulans (FGSC359) and identified as glyceraldehyde-3-phosphate dehydrogenase (gpdA) on the basis of its nucleotide sequence. The level of gpdA RNA substantially increased in cultures gradually adapted to NaCl but was greatly reduced in cultures exposed briefly to a high concentration of NaCl. A pyrG auxotroph of A. nidulans (A773) was cotransformed with a gpdA-uidAconstruct and a plasmid containing the Neurospora crassa pyr4 gene as a selectable marker. OnepyrG + β-glucuronidase-positive (GUS+) transformant was selected, and stable integration of the gpdA-uidA construct into the genome was confirmed by Southern blot analysis. Gradual adaptation to increasing concentrations of NaCl resulted in an increase in GUS activity to 2.7-fold. GUS activity was reduced after a 2-h exposure of an unadapted culture to 2 M NaCl but gradually increased to a maximum of twofold after 24 h. GUS activity also increased by 8.4-fold in Na2SO4-adapted cultures, 4.9-fold in polyethylene glycol-adapted cultures, and 7.5-fold in KCl-adapted cultures. These results are consistent with the hypothesis that theA. nidulans gpdA promoter is transcriptionally activated by osmotic signals.