Analysis of the 3-phosphoglycerate kinase 2 promoter in Rhizopus niveus.
Promoter analysis was performed on the Rhizopus niveus 3-phosphoglycerate kinase 2-encoding gene (pgk2), one of the two pgk genes (pgk1 and pgk2) from this filamentous fungus sequenced so far. Deletion mutants of the promoter region were fused to the Escherichia coli uidA gene (which codes for beta-glucuronidase; GUS), and introduced into R. niveus to measure the intracellular GUS activities of the transformants. Deletion of the sequence between nt -174 to -133 (numbers indicate the position from the putative translation start codon) caused a significant decrease in the ratio of the GUS activity of the transformant cultured in glucose medium compared to that in glycerol medium. In this region, a 21-nt sequence which is well conserved between pgk1 and pgk2 is present. When it was inserted into the promoter region of the uninducible gene encoding RNase Rh of R. niveus, ligated in front of uidA and introduced into R. niveus, the GUS activity of the transformant was greatly induced by glucose, but less by glycerol. We therefore suggest that the 21-nt sequence is a glucose-inducible transcriptional activator of R. niveus. This is the first report on a transcriptional activator in zygomycetes.
Takaya
N
N
Department of Agricultural Chemistry, University of Tokyo, Japan.
Yanai
K
K
Horiuchi
H
H
Ohta
A
A
Takagi
M
M
D001483:Base Sequence; D004252:DNA Mutational Analysis; D004790:Enzyme Induction; D015966:Gene Expression Regulation, Fungal / Q000187:drug effects; D005800:Genes, Fungal / Q000235:genetics*; D005947:Glucose / Q000494:pharmacology; D005966:Glucuronidase / Q000096:biosynthesis / Q000235:genetics; D008969:Molecular Sequence Data; D010735:Phosphoglycerate Kinase / Q000096:biosynthesis / Q000235:genetics*; D011401:Promoter Regions, Genetic / Q000235:genetics*; D011993:Recombinant Fusion Proteins; D012233:Rhizopus / Q000235:genetics*; D017384:Sequence Deletion
D011993:Recombinant Fusion Proteins; D010735:Phosphoglycerate Kinase; D005966:Glucuronidase; D005947:Glucose