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Genetics and Molecular Biology

The Transcriptional Activator XlnR Regulates Both Xylanolytic and Endoglucanase Gene Expression inAspergillus niger

Noël N. M. E. van Peij, Marco M. C. Gielkens, Ronald P. de Vries, Jaap Visser, Leo H. de Graaff
Noël N. M. E. van Peij
Section Molecular Genetics of Industrial Microorganisms, Wageningen Agricultural University, NL-6703 HA Wageningen, The Netherlands
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Marco M. C. Gielkens
Section Molecular Genetics of Industrial Microorganisms, Wageningen Agricultural University, NL-6703 HA Wageningen, The Netherlands
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Ronald P. de Vries
Section Molecular Genetics of Industrial Microorganisms, Wageningen Agricultural University, NL-6703 HA Wageningen, The Netherlands
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Jaap Visser
Section Molecular Genetics of Industrial Microorganisms, Wageningen Agricultural University, NL-6703 HA Wageningen, The Netherlands
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Leo H. de Graaff
Section Molecular Genetics of Industrial Microorganisms, Wageningen Agricultural University, NL-6703 HA Wageningen, The Netherlands
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DOI: 10.1128/AEM.64.10.3615-3619.1998
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    Fig. 1.

    Northern blot analysis of expression of A. niger genes encoding cellulose- and xylan-degrading enzymes. (A) Time course of induction in A. nigerNW205::130 (wt) and NXA1-4 (xlnR1) (a loss-of-function mutant). Both strains were cultured for 18 h in medium containing 3% d-fructose (Fr), and mycelia were subsequently transferred to medium containing 1% xylan (Xa) or 1%d-xylose (Xo) and incubated for the times indicated. Each lane contained 10 μg of total RNA, which was checked by hybridization with the 18S rRNA probe. Blots were hybridized with gene-specific probes as indicated. (B) Comparison of expression of genes encoding cellulose- and xylan-degrading enzymes in A. niger N902 (wt), NXA1-4 (xlnR1), and N902-pIM230-25.12 (XlnR+) (N902 with multiple copies of xlnR) upon transfer to medium containing 1% d-xylose for 6 h after growth for 18 h in medium containing 1%d-fructose. A Northern blot analysis was performed exactly as described above. The signal intensities of the different blots cannot be compared to each other due to the unknown specific activities of the probes used and the different exposure times used for the various blots.

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  • Table 1.

    Probes used in Northern blot analysis

    GeneEMBL accession no.Enzyme encodedFragment usedReference
    abfBX74777α-l-Arabinofuranosidase B1.7-kb EcoRI-XhoI15
    aguAY15405α-Glucuronidase A0.8-kbEcoRV-KpnIa12
    axeAA22880Acetylxylan esterase A1.5-kbHindIIIa,b9
    axhAZ78011Arabinoxylan hydrolase A1.2-kbEcoRI-XhoI17
    bglAβ-Glucosidase A1.0-kbNcoI-SstIThis study
    eglAAJ224451Endoglucanase A0.9-kbXhoIThis study
    eglBAJ224452Endoglucanase B1.1-kbEcoRI-XhoIThis study
    faeAY09330Feruloyl esterase A0.5-kbEcoRV-XhoIa11
    xlnBD38071Endoxylanase B0.9-kbEcoRI-XhoI24
    xlnCEndoxylanase C1.2-kbEcoRI-XhoI18
    xlnDZ84377β-Xylosidase D2.8-kbPstI-NsiIa43
    18S rRNAX7853818S rRNA subunit0.7-kbEcoRI33
    • ↵a Genomic fragment instead of cDNA.

    • ↵b Fragment from the Aspergillus tubingensis axeA gene.

  • Table 2.

    Putative XlnR binding sites in the upstream region of XlnR-controlled genes

    GeneXlnR binding sitePosition(s)a
    aguAGGCTAAa−276
    axeAGGCTAAt−261 (R)
    axhAGGCTAAt−340
    GGCTAAg−850 (R)
    eglAGGCTAAg−710
    eglBGGCTAAg−128
    faeAGGCTAAa−265, −225
    xlnBGGCTAAa−124, −216
    xlnCGGCTAAt−290
    GGCTAAg−500
    xlnDGGCTAAa−133, −147
    • ↵a Position relative to the ATG translation start codon. (R) indicates the opposite orientation of the putative XlnR binding site.

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The Transcriptional Activator XlnR Regulates Both Xylanolytic and Endoglucanase Gene Expression inAspergillus niger
Noël N. M. E. van Peij, Marco M. C. Gielkens, Ronald P. de Vries, Jaap Visser, Leo H. de Graaff
Applied and Environmental Microbiology Oct 1998, 64 (10) 3615-3619; DOI: 10.1128/AEM.64.10.3615-3619.1998

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The Transcriptional Activator XlnR Regulates Both Xylanolytic and Endoglucanase Gene Expression inAspergillus niger
Noël N. M. E. van Peij, Marco M. C. Gielkens, Ronald P. de Vries, Jaap Visser, Leo H. de Graaff
Applied and Environmental Microbiology Oct 1998, 64 (10) 3615-3619; DOI: 10.1128/AEM.64.10.3615-3619.1998
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KEYWORDS

Aspergillus niger
cellulase
Fungal Proteins
Gene Expression Regulation, Fungal
Glucosidases
Trans-Activators
Transcription, Genetic

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