ACEII, a Novel Transcriptional Activator Involved in Regulation of Cellulase and Xvlanase Genes of Trichoderma reesei

Research output: Contribution to journalArticleScientificpeer-review

212 Citations (Scopus)

Abstract

A novel yeast-based method to isolate transcriptional activators was applied to clone regulators binding to the cellulase promoter cbh1 of the filamentous fungus Trichoderma reesei (Hypocrea jecorina). This led to the isolation of the cellulase activator ace2 encoding for a protein belonging to the class of zinc binuclear cluster proteins found exclusively in fungi. The DNA-binding domain of ACEII was expressed as a glutathione S-transferase fusion protein in Escherichia coli, and ACEII was shown to bind in vitro to the 5′-GGCTAATAA site present in the cbh1 promoter. This site also contains the proposed binding sequence of the xylanase activator XlnR of Aspergillus niger. Mutation of the GGC triplet abolished ACEII binding. The function of ACEII was studied by analyzing the effects of ace2 deletion in the hypercellulolytic T. reesei strain ALKO2221. Deletion of the ace2 gene led to lowered induction kinetics of mRNAs encoding the major cellulases cellobiohydrolases I and II and endoglucanases I and II and to 30-70% reduced cellulase activity when the fungus was grown on medium containing Solka floc cellulose. The expression level of the gene encoding xylanase was also affected. ace2 deletion led to lowered xyn2 expression in cellulose-induced cultivation. Cellulase induction by sophorose was not affected by ace2 deletion.

Original languageEnglish
Pages (from-to)24309-24314
Number of pages6
JournalJournal of Biological Chemistry
Volume276
Issue number26
DOIs
Publication statusPublished - 29 Jun 2001
MoE publication typeA1 Journal article-refereed

Fingerprint

Trichoderma
Cellulase
Genes
Fungi
Cellulose 1,4-beta-Cellobiosidase
Cellulases
Cellulose
Hypocrea
Proteins
Gene encoding
Aspergillus niger
Aspergillus
Escherichia coli Proteins
Gene Deletion
Glutathione Transferase
Yeast
Escherichia coli
Zinc
Fusion reactions
Clone Cells

Keywords

  • cells
  • enzyme kinetics
  • Enzymes
  • genes
  • Yeast
  • gene coding
  • ACEII protein
  • cellulase
  • Aspergillus
  • Aspergillus niger
  • Escherichia coli
  • Fungi
  • Hypocrea jecorina
  • Trichoderma

Cite this

@article{1d6294ef2a6a47e8944e05a14c407741,
title = "ACEII, a Novel Transcriptional Activator Involved in Regulation of Cellulase and Xvlanase Genes of Trichoderma reesei",
abstract = "A novel yeast-based method to isolate transcriptional activators was applied to clone regulators binding to the cellulase promoter cbh1 of the filamentous fungus Trichoderma reesei (Hypocrea jecorina). This led to the isolation of the cellulase activator ace2 encoding for a protein belonging to the class of zinc binuclear cluster proteins found exclusively in fungi. The DNA-binding domain of ACEII was expressed as a glutathione S-transferase fusion protein in Escherichia coli, and ACEII was shown to bind in vitro to the 5′-GGCTAATAA site present in the cbh1 promoter. This site also contains the proposed binding sequence of the xylanase activator XlnR of Aspergillus niger. Mutation of the GGC triplet abolished ACEII binding. The function of ACEII was studied by analyzing the effects of ace2 deletion in the hypercellulolytic T. reesei strain ALKO2221. Deletion of the ace2 gene led to lowered induction kinetics of mRNAs encoding the major cellulases cellobiohydrolases I and II and endoglucanases I and II and to 30-70{\%} reduced cellulase activity when the fungus was grown on medium containing Solka floc cellulose. The expression level of the gene encoding xylanase was also affected. ace2 deletion led to lowered xyn2 expression in cellulose-induced cultivation. Cellulase induction by sophorose was not affected by ace2 deletion.",
keywords = "cells, enzyme kinetics, Enzymes, genes, Yeast, gene coding, ACEII protein, cellulase, Aspergillus, Aspergillus niger, Escherichia coli, Fungi, Hypocrea jecorina, Trichoderma",
author = "Nina Aro and Anu Saloheimo and Marja Ilm{\'e}n and Merja Penttil{\"a}",
year = "2001",
month = "6",
day = "29",
doi = "10.1074/jbc.M003624200",
language = "English",
volume = "276",
pages = "24309--24314",
journal = "Journal of Biological Chemistry",
issn = "0021-9258",
publisher = "American Society for Biochemistry and Molecular Biology Inc.",
number = "26",

}

ACEII, a Novel Transcriptional Activator Involved in Regulation of Cellulase and Xvlanase Genes of Trichoderma reesei. / Aro, Nina; Saloheimo, Anu; Ilmén, Marja; Penttilä, Merja.

In: Journal of Biological Chemistry, Vol. 276, No. 26, 29.06.2001, p. 24309-24314.

Research output: Contribution to journalArticleScientificpeer-review

TY - JOUR

T1 - ACEII, a Novel Transcriptional Activator Involved in Regulation of Cellulase and Xvlanase Genes of Trichoderma reesei

AU - Aro, Nina

AU - Saloheimo, Anu

AU - Ilmén, Marja

AU - Penttilä, Merja

PY - 2001/6/29

Y1 - 2001/6/29

N2 - A novel yeast-based method to isolate transcriptional activators was applied to clone regulators binding to the cellulase promoter cbh1 of the filamentous fungus Trichoderma reesei (Hypocrea jecorina). This led to the isolation of the cellulase activator ace2 encoding for a protein belonging to the class of zinc binuclear cluster proteins found exclusively in fungi. The DNA-binding domain of ACEII was expressed as a glutathione S-transferase fusion protein in Escherichia coli, and ACEII was shown to bind in vitro to the 5′-GGCTAATAA site present in the cbh1 promoter. This site also contains the proposed binding sequence of the xylanase activator XlnR of Aspergillus niger. Mutation of the GGC triplet abolished ACEII binding. The function of ACEII was studied by analyzing the effects of ace2 deletion in the hypercellulolytic T. reesei strain ALKO2221. Deletion of the ace2 gene led to lowered induction kinetics of mRNAs encoding the major cellulases cellobiohydrolases I and II and endoglucanases I and II and to 30-70% reduced cellulase activity when the fungus was grown on medium containing Solka floc cellulose. The expression level of the gene encoding xylanase was also affected. ace2 deletion led to lowered xyn2 expression in cellulose-induced cultivation. Cellulase induction by sophorose was not affected by ace2 deletion.

AB - A novel yeast-based method to isolate transcriptional activators was applied to clone regulators binding to the cellulase promoter cbh1 of the filamentous fungus Trichoderma reesei (Hypocrea jecorina). This led to the isolation of the cellulase activator ace2 encoding for a protein belonging to the class of zinc binuclear cluster proteins found exclusively in fungi. The DNA-binding domain of ACEII was expressed as a glutathione S-transferase fusion protein in Escherichia coli, and ACEII was shown to bind in vitro to the 5′-GGCTAATAA site present in the cbh1 promoter. This site also contains the proposed binding sequence of the xylanase activator XlnR of Aspergillus niger. Mutation of the GGC triplet abolished ACEII binding. The function of ACEII was studied by analyzing the effects of ace2 deletion in the hypercellulolytic T. reesei strain ALKO2221. Deletion of the ace2 gene led to lowered induction kinetics of mRNAs encoding the major cellulases cellobiohydrolases I and II and endoglucanases I and II and to 30-70% reduced cellulase activity when the fungus was grown on medium containing Solka floc cellulose. The expression level of the gene encoding xylanase was also affected. ace2 deletion led to lowered xyn2 expression in cellulose-induced cultivation. Cellulase induction by sophorose was not affected by ace2 deletion.

KW - cells

KW - enzyme kinetics

KW - Enzymes

KW - genes

KW - Yeast

KW - gene coding

KW - ACEII protein

KW - cellulase

KW - Aspergillus

KW - Aspergillus niger

KW - Escherichia coli

KW - Fungi

KW - Hypocrea jecorina

KW - Trichoderma

UR - http://www.scopus.com/inward/record.url?scp=0035968258&partnerID=8YFLogxK

U2 - 10.1074/jbc.M003624200

DO - 10.1074/jbc.M003624200

M3 - Article

VL - 276

SP - 24309

EP - 24314

JO - Journal of Biological Chemistry

JF - Journal of Biological Chemistry

SN - 0021-9258

IS - 26

ER -