Different inducibility of expression of the two xylanase genes xyn1 and xyn2 in Trichoderma reesei

J Biol Chem. 1996 Oct 11;271(41):25624-9. doi: 10.1074/jbc.271.41.25624.

Abstract

Regulation of formation of the extracellular xylanase system of Trichoderma reesei QM 9414 during growth on xylan, cellulose, and replacement onto a number of soluble inducers was investigated by Northern analysis of xyn1 and xyn2 transcripts and by the use of the Escherichia coli hph (hygromycin B-phosphotransferase-encoding) gene as a reporter. Whereas the xyn1 promoter is active in the presence of xylan and xylose, and virtually silenced in the presence of glucose, the xyn2 promoter enables basal transcription at a low level, but is enhanced in the presence of xylan and xylobiose and also of sophorose or cellobiose. The respective regulatory nucleotide regions were localized on a 221-base pair fragment and a 55-base pair fragment of the xyn1 and xyn2 5'-upstream noncoding sequences, respectively. Electrophoretic mobility shift assays, using cell-free extracts, identified induction-specific protein-DNA complexes: one complex of high mobility was observed under basal, noninduced conditions (glucose) with xyn2, which was in part replaced by a slow-migrating complex upon induction by xylan or sophorose. Both complexes bound to a CCAAT box. With xyn1, the induced complex also binds to a CCAAT box, but this binding is not observed in the presence of the carbon catabolite repressor Cre1, which binds to a nearby located consensus motif.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Base Sequence
  • Carbohydrates / pharmacology
  • DNA Primers
  • Enzyme Induction / drug effects
  • Gene Expression Regulation, Fungal* / drug effects
  • Genes, Reporter
  • Molecular Sequence Data
  • Polymerase Chain Reaction
  • Promoter Regions, Genetic
  • Recombinant Proteins / biosynthesis
  • Transcription, Genetic*
  • Trichoderma / enzymology*
  • Trichoderma / genetics*
  • Trichoderma / growth & development
  • Xylan Endo-1,3-beta-Xylosidase
  • Xylosidases / biosynthesis*
  • Xylosidases / genetics

Substances

  • Carbohydrates
  • DNA Primers
  • Recombinant Proteins
  • Xylosidases
  • Xylan Endo-1,3-beta-Xylosidase