Alkaline pH-induced up-regulation of the afp gene encoding the antifungal protein (AFP) of Aspergillus giganteus is not mediated by the transcription factor PacC: possible involvement of calcineurin.
The afp gene encoding the antifungal protein (AFP) of Aspergillus giganteus has a prototypical alkaline gene expression pattern, which suggests that the gene might be under the control of the ambient pH-dependent zinc-finger transcription factor PacC. This notion is corroborated by the presence in the upstream region of afp of two putative PacC binding sites, afpP1 and afpP2, which are specifically recognised by the PacC protein of A. nidulans in vitro. However, in this report we provide several lines of evidence to show that pH-dependent up-regulation of afp is not mediated by transcriptional activation through PacC. (1) The temporal expression pattern of the A. giganteus pacC gene does not parallel the accumulation of the afp mRNA during cultivation. (2) Inactivation of afpP1 and afpP2 did not reduce promoter activity under alkaline conditions, as determined from the relative wild-type and mutant afp::lacZ reporter activities in A. nidulans. (3) Reporter activities in acidity- and alkalinity-mimicking mutant strains are inconsistent with a positive role for PacC in afp expression. (4) In A. giganteus, the pH-dependent increase in afp mRNA and AFP levels can be completely prevented by the calcineurin inhibitor FK506, suggesting that the calcineurin signalling pathway might control the in vivo activation of the afp promoter by alkaline pH.
Meyer
Vera
V
Institut für Biotechnologie, Fachgebiet Mikrobiologie und Genetik, Technische Universität Berlin, Germany. v.meyer@lb.tu-berlin.de
Spielvogel
Anja
A
Funk
Laura
L
Tilburn
Joan
J
Arst
Herbert N
HN
Stahl
Ulf
U
D000595:Amino Acid Sequence; D001230:Aspergillus / Q000235:genetics* / Q000378:metabolism; D001483:Base Sequence; D019703:Calcineurin / Q000378:metabolism; D065095:Calcineurin Inhibitors; D017931:DNA Primers; D005656:Fungal Proteins / Q000378:metabolism*; D006863:Hydrogen-Ion Concentration; D008969:Molecular Sequence Data; D009154:Mutation / Q000235:genetics; D011401:Promoter Regions, Genetic / Q000187:drug effects / Q000235:genetics; D020133:Reverse Transcriptase Polymerase Chain Reaction; D017422:Sequence Analysis, DNA; D016559:Tacrolimus / Q000494:pharmacology; D014157:Transcription Factors / Q000378:metabolism*; D015854:Up-Regulation / Q000187:drug effects* / Q000235:genetics*
D065095:Calcineurin Inhibitors; D017931:DNA Primers; D005656:Fungal Proteins; C092394:PacC protein, Aspergillus; D014157:Transcription Factors; C097609:antifungal protein, Aspergillus; D019703:Calcineurin; D016559:Tacrolimus