Deletion analysis of the enolase gene (enoA) promoter from the filamentous fungus Aspegillus oryzae.
The enolase gene (enoA) is one of the most strongly expressed genes in Aspergillus oryzae. To elucidate the transcription regulatory element for this strong expression and the process of glucose induction, the transcription activity of a series of truncated enoA promoters was measured by using the Escherichia coli uidA gene as a reporter. Deletion of a 104-bp region located -224 nt to -121 nt upstream of the translation initiation site caused both a drastic decrease in the beta-glucuronidase (GUS) activity and a loss of glucose induction. Northern blot analysis confirmed that the decrease in GUS activity was achieved at the transcriptional level. In addition, electrophoretic gel mobility shift assays indicated that the 104-bp region contained a 15-bp element, to which one or more A. oryzae cellular factors specifically bind. These results suggest that the 15-bp element between -195 nt and -181 nt includes the sequence essential for the transcription regulation of the A. oryzae enoA gene.
Toda
T
T
Institute of Molecular and Cell Biology, National Institute of Advanced Industrial Science and Technology, Tsukuba, Ibaraki, Japan.
Sano
M
M
Honda
M
M
Rimoldi
O J
OJ
Yang
Y
Y
Yamamoto
M
M
Takase
K
K
Hirozumi
K
K
Kitamoto
K
K
Minetoki
T
T
Gomi
K
K
Machida
M
M
D001236:Aspergillus oryzae / Q000201:enzymology / Q000235:genetics*; D001483:Base Sequence; D004252:DNA Mutational Analysis; D017353:Gene Deletion; D015966:Gene Expression Regulation, Fungal; D005800:Genes, Fungal / Q000235:genetics*; D005966:Glucuronidase / Q000378:metabolism; D008969:Molecular Sequence Data; D010751:Phosphopyruvate Hydratase / Q000235:genetics*; D011401:Promoter Regions, Genetic / Q000235:genetics*
D005966:Glucuronidase; D010751:Phosphopyruvate Hydratase