Functional analysis of the endoxylanase B (xynB) promoter from Penicillium purpurogenum.
In Penicillium purpurogenum, the gene encoding endoxylanase B (xynB) is highly expressed by xylan and repressed by glucose at the transcriptional level. The promoter of this gene has a modular structure, with eight putative XlnR binding sites in tandem (XlnR module), and upstream from them, four putative CreA binding sites (CreA module). Promoter fragments containing different modules were inserted into a plasmid, pAN49-1, which contains a basal fungal promoter linked to a reporter gene (lacZ) and its expression was studied in vivo in Aspergillus nidulans. The XlnR module is able to trigger high beta-galactosidase activity in the presence of xylan, but the lack of most XlnR sites notoriously reduces this enzymatic activity. No enzyme induction is observed if the orientation of the promoter fragment is inverted. The presence of the CreA module is necessary for glucose repression when beta-galactosidase activity is previously induced by xylan. However, when transformant strains containing the XlnR module but lacking all CreA sites were grown in glucose without pre-induction in xylan, a low beta-galactosidase activity was observed compared with the same transformants grown in xylan. These results agree with a double-lock regulatory mechanism for both direct and indirect repression of xylanolytic genes by glucose.
Díaz
Jheimmy
J
Departamento de Genética Molecular y Microbiología, Facultad de Ciencias Biológicas, P. Universidad Católica de Chile, Alameda 340, Casilla 114-D, Santiago, Chile.
Chávez
Renato
R
Larrondo
Luis F
LF
Eyzaguirre
Jaime
J
Bull
Paulina
P
D001230:Aspergillus / Q000235:genetics / Q000254:growth & development; D001483:Base Sequence; D015139:Blotting, Southern; D017931:DNA Primers; D043364:Endo-1,4-beta Xylanases / Q000235:genetics*; D005800:Genes, Fungal; D017930:Genes, Reporter; D005966:Glucuronidase / Q000235:genetics; D010407:Penicillium / Q000235:genetics*; D010957:Plasmids; D011401:Promoter Regions, Genetic; D014170:Transformation, Genetic
D017931:DNA Primers; D005966:Glucuronidase; D043364:Endo-1,4-beta Xylanases