Regulation of the alpha-glucuronidase-encoding gene ( aguA) from Aspergillus niger.
The alpha-glucuronidase gene aguA from Aspergillus niger was cloned and characterised. Analysis of the promoter region of aguA revealed the presence of four putative binding sites for the major carbon catabolite repressor protein CREA and one putative binding site for the transcriptional activator XLNR. In addition, a sequence motif was detected which differed only in the last nucleotide from the XLNR consensus site. A construct in which part of the aguA coding region was deleted still resulted in production of a stable mRNA upon transformation of A. niger. The putative XLNR binding sites and two of the putative CREA binding sites were mutated individually in this construct and the effects on expression were examined in A. niger transformants. Northern analysis of the transformants revealed that the consensus XLNR site is not actually functional in the aguA promoter, whereas the sequence that diverges from the consensus at a single position is functional. This indicates that XLNR is also able to bind to the sequence GGCTAG, and the XLNR binding site consensus should therefore be changed to GGCTAR. Both CREA sites are functional, indicating that CREA has a strong influence on aguA expression. A detailed expression analysis of aguA in four genetic backgrounds revealed a second regulatory system involved in activation of aguA gene expression. This system responds to the presence of glucuronic and galacturonic acids, and is not dependent on XLNR.
de Vries
R P
RP
Department of Microbiology, Utrecht University, Padualaan 8, 3508 TB Utrecht, The Netherlands. r.p.devries@bio.uu.nl
van de Vondervoort
P J I
PJ
Hendriks
L
L
van de Belt
M
M
Visser
J
J
D001234:Aspergillus niger / Q000187:drug effects / Q000201:enzymology / Q000235:genetics*; D001665:Binding Sites; D003001:Cloning, Molecular; D005656:Fungal Proteins / Q000378:metabolism; D015971:Gene Expression Regulation, Enzymologic / Q000187:drug effects; D017930:Genes, Reporter; D005818:Genetic Engineering; D006026:Glycoside Hydrolases / Q000235:genetics*; D008969:Molecular Sequence Data; D009154:Mutation / Q000235:genetics; D011401:Promoter Regions, Genetic / Q000235:genetics; D012097:Repressor Proteins / Q000378:metabolism; D015534:Trans-Activators / Q000378:metabolism; D014994:Xylose / Q000494:pharmacology
D005656:Fungal Proteins; D012097:Repressor Proteins; D015534:Trans-Activators; C115443:XlnR protein, Aspergillus; C071237:CreA protein, Aspergillus nidulans; D014994:Xylose; D006026:Glycoside Hydrolases; C096734:alpha-glucuronidase