Identification of the cis-acting elements involved in regulation of xylanase III gene expression in Trichoderma reesei PC-3-7.
The xylanase III gene (xyn3) from the filamentous fungus Trichoderma reesei PC-3-7 is only induced by cellulose, its derivatives and L-sorbose, but not by xylan. In this study, we defined three cis-acting elements within the xyn3 upstream region by using detailed deletion and mutation analysis. In addition to the Xyr1/ACEII-binding motif (5'-GGCTAA-3'), the analogous motifs 5'-GGCTAT-3' and 5'-GGCAAA-3' presented as an inverted repeat spaced by 16-bp internal sequences were identified as essential elements for xyn3 expression. Electrophoretic mobility shift assay using heterologously expressed Xyr1 demonstrates that all the identified cis-acting elements are able to interact with Xyr1. Furthermore, no xyn3 transcripts were formed in the xyr1-knockout strain upon induction by sophorose and L-sorbose. These results indicate that xyn3 expression is transcriptionally regulated by Xyr1, and suggest that the 5'-GGCTAT-3' and 5'-GGCAAA-3' motifs play roles in Xyr1-mediated cellulase and xylanase gene expression in T. reesei.
Furukawa
Takanori
T
Department of Bioengineering, Nagaoka University of Technology, 1603-1, Kamitomioka, Nagaoka, Niigata 940-2188, Japan.
Shida
Yosuke
Y
Kitagami
Naoki
N
Ota
Yuuki
Y
Adachi
Masahiro
M
Nakagawa
Shiho
S
Shimada
Ryuichi
R
Kato
Masashi
M
Kobayashi
Tetsuo
T
Okada
Hirofumi
H
Ogasawara
Wataru
W
Morikawa
Yasushi
Y
D001483:Base Sequence; D004268:DNA-Binding Proteins / Q000737:chemistry / Q000235:genetics / Q000378:metabolism; D024202:Electrophoretic Mobility Shift Assay; D005656:Fungal Proteins / Q000737:chemistry / Q000235:genetics / Q000378:metabolism; D015966:Gene Expression Regulation, Fungal; D017930:Genes, Reporter; D020218:Response Elements; D017384:Sequence Deletion; D014158:Transcription, Genetic; D014242:Trichoderma / Q000201:enzymology* / Q000235:genetics* / Q000378:metabolism; D014995:Xylosidases / Q000235:genetics* / Q000378:metabolism
D004268:DNA-Binding Proteins; D005656:Fungal Proteins; D014995:Xylosidases