Novel reporter gene expression systems for monitoring activation of the Aspergillus nidulans HOG pathway.
The Aspergillus nidulans high-osmolarity glycerol response (AnHOG) pathway is involved in osmoadaptation. We found that fludioxonil, a fungicide, causes improper activation of HogA mitogen-activated protein kinase (MAPK) in A. nidulans. Here we present novel reporter systems for monitoring activation of the AnHOG pathway. The promoter region of gfdB (glycerol-3-phosphate dehydrogenase), whose expression depends on the presence of HogA, was fused to a beta-glucuronidase uidA gene (GUS) to construct the reporter, which was introduced into A. nidulans wild type and hogADelta. Increased GUS activity was detected in the wild type only when it was treated with high osmolarity or fludioxonil, while reporter activity was scarcely stimulated in the hogADelta mutant. These results indicate that the reporter activity is controlled via HogA activation. Furthermore, we present possible applications of the reporter systems in screening new antifungal compounds.
Furukawa
Kentaro
K
Laboratory of Enzymology, Graduate School of Agricultural Science, Tohoku University, Sendai, Japan. kabe@biochem.tohoku.ac.jp
Yoshimi
Akira
A
Furukawa
Takako
T
Hoshi
Yukiko
Y
Hagiwara
Daisuke
D
Sato
Natsuko
N
Fujioka
Tomonori
T
Mizutani
Osamu
O
Mizuno
Takeshi
T
Kobayashi
Tetsuo
T
Abe
Keietsu
K
D001233:Aspergillus nidulans / Q000187:drug effects* / Q000201:enzymology / Q000254:growth & development; D004149:Dioxoles / Q000494:pharmacology*; D004789:Enzyme Activation / Q000187:drug effects; D015966:Gene Expression Regulation, Fungal / Q000187:drug effects*; D018014:Gene Transfer Techniques; D017930:Genes, Reporter; D005990:Glycerol / Q000378:metabolism*; D020935:MAP Kinase Signaling System / Q000187:drug effects*; D011758:Pyrroles / Q000494:pharmacology*
D004149:Dioxoles; D011758:Pyrroles; C108339:fludioxonil; D005990:Glycerol