Proteome and transcriptome analysis of the cellular responses to production of the heterologous protein, Melanocarpus albomyces laccase, in the filamentous fungus Trichoderma reesei

Tiina Pakula (Corresponding author), Jari Rautio, Bart Smit, Mikko Arvas, Markku Saloheimo, Merja Penttilä

Research output: Chapter in Book/Report/Conference proceedingConference abstract in proceedingsScientific

Abstract

Trichoderma reesei Rut-C30 and its transformant producing Melanocarpus
albomyces
laccase were cultivated in lactose-containing minimal medium in
chemostats and subjected to proteome and transcriptome analysis to identify
differentially expressed genes under these conditions. The cultures were carried
out using a dilution rate (0.03/h) previously shown to be optimal for production
of endogenous extracellular proteins under these conditions. The TRAC method
for multiplexed transcriptional analysis was first applied to analyse the
expression levels of a selected set of genes and to monitor the steady state in
the chemostat cultures. The results indicated that UPR (unfolded protein
response) pathway was not activated as a response to laccase production. On
the contrary, the expression levels of e.g. the foldase and chaperone genes pdi1
and bip1 were expressed at a slightly higher level in the host. Also the
expression levels of the genes encoding the endogenous secreted cellulases,
egl1 and cbh1, were slightly higher in the host strain. Both proteome and
genome wide transcriptional analysis using oligonucleotide microarrays revealed
only a modest number of cellular responses to laccase production. Altogether 32
genes were differentially expressed between the strains, only five of the genes
being upregulated in the laccase producing strain, and the rest having a higher
expression level in the host strain and including many genes encoding
extracellular enzymes. Proteome analysis using 2D gel electrophoresis with DIGE
showed increased expression of a group of proteins including e.g. heat shock
proteins as well ubiquitin associated proteins and proteins involved in ERassociated protein degradation. The result indicates that although laccase
expression does not induce apparent UPR, other type stress responses are
induced e.g. activated protein degradation. The results were also compared to
those obtained in previous studies on the production human tissue plasminogen
activator (tPA) in T. reesei.
Original languageEnglish
Title of host publication8th European Conference on Fungal Genetics
Subtitle of host publicationBook of Abstracts
EditorsIrina S. Druzhnina, Alexey G. Kopchinskiy, Christian P. Kubicek
PublisherVienna University of Technology
ChapterVII Cell-Factories and Biotechnology
Pages290
Publication statusPublished - 2006
Event8th European Conference on Fungal Genetics - Vienna, Austria
Duration: 8 Apr 200611 Apr 2006

Conference

Conference8th European Conference on Fungal Genetics
CountryAustria
CityVienna
Period8/04/0611/04/06

Fingerprint

Trichoderma reesei
laccase
proteome
transcriptomics
fungi
host strains
genes
proteins
cellulases
ubiquitin
oligonucleotides
protein degradation
gel electrophoresis
lactose
stress response
heterologous gene expression
heat
degradation
enzymes

Cite this

Pakula, T., Rautio, J., Smit, B., Arvas, M., Saloheimo, M., & Penttilä, M. (2006). Proteome and transcriptome analysis of the cellular responses to production of the heterologous protein, Melanocarpus albomyces laccase, in the filamentous fungus Trichoderma reesei. In I. S. Druzhnina, A. G. Kopchinskiy, & C. P. Kubicek (Eds.), 8th European Conference on Fungal Genetics: Book of Abstracts (pp. 290). [VIIp-35] Vienna University of Technology.
Pakula, Tiina ; Rautio, Jari ; Smit, Bart ; Arvas, Mikko ; Saloheimo, Markku ; Penttilä, Merja. / Proteome and transcriptome analysis of the cellular responses to production of the heterologous protein, Melanocarpus albomyces laccase, in the filamentous fungus Trichoderma reesei. 8th European Conference on Fungal Genetics: Book of Abstracts. editor / Irina S. Druzhnina ; Alexey G. Kopchinskiy ; Christian P. Kubicek. Vienna University of Technology, 2006. pp. 290
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title = "Proteome and transcriptome analysis of the cellular responses to production of the heterologous protein, Melanocarpus albomyces laccase, in the filamentous fungus Trichoderma reesei",
abstract = "Trichoderma reesei Rut-C30 and its transformant producing Melanocarpusalbomyces laccase were cultivated in lactose-containing minimal medium inchemostats and subjected to proteome and transcriptome analysis to identifydifferentially expressed genes under these conditions. The cultures were carriedout using a dilution rate (0.03/h) previously shown to be optimal for productionof endogenous extracellular proteins under these conditions. The TRAC methodfor multiplexed transcriptional analysis was first applied to analyse theexpression levels of a selected set of genes and to monitor the steady state inthe chemostat cultures. The results indicated that UPR (unfolded proteinresponse) pathway was not activated as a response to laccase production. Onthe contrary, the expression levels of e.g. the foldase and chaperone genes pdi1and bip1 were expressed at a slightly higher level in the host. Also theexpression levels of the genes encoding the endogenous secreted cellulases,egl1 and cbh1, were slightly higher in the host strain. Both proteome andgenome wide transcriptional analysis using oligonucleotide microarrays revealedonly a modest number of cellular responses to laccase production. Altogether 32genes were differentially expressed between the strains, only five of the genesbeing upregulated in the laccase producing strain, and the rest having a higherexpression level in the host strain and including many genes encodingextracellular enzymes. Proteome analysis using 2D gel electrophoresis with DIGEshowed increased expression of a group of proteins including e.g. heat shockproteins as well ubiquitin associated proteins and proteins involved in ERassociated protein degradation. The result indicates that although laccaseexpression does not induce apparent UPR, other type stress responses areinduced e.g. activated protein degradation. The results were also compared tothose obtained in previous studies on the production human tissue plasminogenactivator (tPA) in T. reesei.",
author = "Tiina Pakula and Jari Rautio and Bart Smit and Mikko Arvas and Markku Saloheimo and Merja Penttil{\"a}",
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language = "English",
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editor = "Druzhnina, {Irina S.} and Kopchinskiy, {Alexey G.} and Kubicek, {Christian P.}",
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Pakula, T, Rautio, J, Smit, B, Arvas, M, Saloheimo, M & Penttilä, M 2006, Proteome and transcriptome analysis of the cellular responses to production of the heterologous protein, Melanocarpus albomyces laccase, in the filamentous fungus Trichoderma reesei. in IS Druzhnina, AG Kopchinskiy & CP Kubicek (eds), 8th European Conference on Fungal Genetics: Book of Abstracts., VIIp-35, Vienna University of Technology, pp. 290, 8th European Conference on Fungal Genetics, Vienna, Austria, 8/04/06.

Proteome and transcriptome analysis of the cellular responses to production of the heterologous protein, Melanocarpus albomyces laccase, in the filamentous fungus Trichoderma reesei. / Pakula, Tiina (Corresponding author); Rautio, Jari; Smit, Bart; Arvas, Mikko; Saloheimo, Markku; Penttilä, Merja.

8th European Conference on Fungal Genetics: Book of Abstracts. ed. / Irina S. Druzhnina; Alexey G. Kopchinskiy; Christian P. Kubicek. Vienna University of Technology, 2006. p. 290 VIIp-35.

Research output: Chapter in Book/Report/Conference proceedingConference abstract in proceedingsScientific

TY - CHAP

T1 - Proteome and transcriptome analysis of the cellular responses to production of the heterologous protein, Melanocarpus albomyces laccase, in the filamentous fungus Trichoderma reesei

AU - Pakula, Tiina

AU - Rautio, Jari

AU - Smit, Bart

AU - Arvas, Mikko

AU - Saloheimo, Markku

AU - Penttilä, Merja

N1 - CA2: TK402

PY - 2006

Y1 - 2006

N2 - Trichoderma reesei Rut-C30 and its transformant producing Melanocarpusalbomyces laccase were cultivated in lactose-containing minimal medium inchemostats and subjected to proteome and transcriptome analysis to identifydifferentially expressed genes under these conditions. The cultures were carriedout using a dilution rate (0.03/h) previously shown to be optimal for productionof endogenous extracellular proteins under these conditions. The TRAC methodfor multiplexed transcriptional analysis was first applied to analyse theexpression levels of a selected set of genes and to monitor the steady state inthe chemostat cultures. The results indicated that UPR (unfolded proteinresponse) pathway was not activated as a response to laccase production. Onthe contrary, the expression levels of e.g. the foldase and chaperone genes pdi1and bip1 were expressed at a slightly higher level in the host. Also theexpression levels of the genes encoding the endogenous secreted cellulases,egl1 and cbh1, were slightly higher in the host strain. Both proteome andgenome wide transcriptional analysis using oligonucleotide microarrays revealedonly a modest number of cellular responses to laccase production. Altogether 32genes were differentially expressed between the strains, only five of the genesbeing upregulated in the laccase producing strain, and the rest having a higherexpression level in the host strain and including many genes encodingextracellular enzymes. Proteome analysis using 2D gel electrophoresis with DIGEshowed increased expression of a group of proteins including e.g. heat shockproteins as well ubiquitin associated proteins and proteins involved in ERassociated protein degradation. The result indicates that although laccaseexpression does not induce apparent UPR, other type stress responses areinduced e.g. activated protein degradation. The results were also compared tothose obtained in previous studies on the production human tissue plasminogenactivator (tPA) in T. reesei.

AB - Trichoderma reesei Rut-C30 and its transformant producing Melanocarpusalbomyces laccase were cultivated in lactose-containing minimal medium inchemostats and subjected to proteome and transcriptome analysis to identifydifferentially expressed genes under these conditions. The cultures were carriedout using a dilution rate (0.03/h) previously shown to be optimal for productionof endogenous extracellular proteins under these conditions. The TRAC methodfor multiplexed transcriptional analysis was first applied to analyse theexpression levels of a selected set of genes and to monitor the steady state inthe chemostat cultures. The results indicated that UPR (unfolded proteinresponse) pathway was not activated as a response to laccase production. Onthe contrary, the expression levels of e.g. the foldase and chaperone genes pdi1and bip1 were expressed at a slightly higher level in the host. Also theexpression levels of the genes encoding the endogenous secreted cellulases,egl1 and cbh1, were slightly higher in the host strain. Both proteome andgenome wide transcriptional analysis using oligonucleotide microarrays revealedonly a modest number of cellular responses to laccase production. Altogether 32genes were differentially expressed between the strains, only five of the genesbeing upregulated in the laccase producing strain, and the rest having a higherexpression level in the host strain and including many genes encodingextracellular enzymes. Proteome analysis using 2D gel electrophoresis with DIGEshowed increased expression of a group of proteins including e.g. heat shockproteins as well ubiquitin associated proteins and proteins involved in ERassociated protein degradation. The result indicates that although laccaseexpression does not induce apparent UPR, other type stress responses areinduced e.g. activated protein degradation. The results were also compared tothose obtained in previous studies on the production human tissue plasminogenactivator (tPA) in T. reesei.

M3 - Conference abstract in proceedings

SP - 290

BT - 8th European Conference on Fungal Genetics

A2 - Druzhnina, Irina S.

A2 - Kopchinskiy, Alexey G.

A2 - Kubicek, Christian P.

PB - Vienna University of Technology

ER -

Pakula T, Rautio J, Smit B, Arvas M, Saloheimo M, Penttilä M. Proteome and transcriptome analysis of the cellular responses to production of the heterologous protein, Melanocarpus albomyces laccase, in the filamentous fungus Trichoderma reesei. In Druzhnina IS, Kopchinskiy AG, Kubicek CP, editors, 8th European Conference on Fungal Genetics: Book of Abstracts. Vienna University of Technology. 2006. p. 290. VIIp-35