Functional studies of the secretory pathway of filamentous fungi

The effect of unfolded protein response on protein production: Dissertation

Research output: ThesisDissertationCollection of Articles

Abstract

This study describes the cloning and characterisation of three genes from the filamentous fungus Trichoderma reesei. Snc1 encoding a putative v-SNARE was shown to be the functional homologue of Saccharomyces cerevisiae SNC1 and SNC2 genes that encode proteins shown to function in exo- and endocytosis. Two components of the unfolded protein response (UPR) pathway have also been identified. Ire1 encodes a serine /threonine kinase that has been shown to be involved the sensing of the protein folding status in the ER and transferring the signal to the nucleus. Ptc2 on the other hand, encodes a putative negative regulator of UPR. Both genes were isolated from T. reesei and shown to be the functional homologues of S. cerevisiae genes. The effects of UPR activation on gene expression and protein production in S. cerevisiae and two filamentous fungi, T. reesei and Aspergillus niger var. awamori are also presented. The data shows that the UPR induction, either by HacA or Ire1 overexpression in A. niger var awamori and T. reesei, respectively, induces the expression of an ER-resident foldase and an ER-resident chaperone. Moreover, the UPR induction induces the expression of genes encoding functions at different steps of the secretory pathway in both fungi. This is in correlation with results obtained in other organisms. The UPR induction was in this study shown to induce production of secreted proteins both in S. cerevisiae and A. niger var. awamori. In yeast the UPR induction by constitutive over-expression of activated yeast HAC1 and T. reesei hac1 resulted in induction of both native and foreign protein production. On the other hand, deletion of HAC1 resulted in decrease in protein production. In A. niger var. awamori only the production of foreign proteins was induced in HacA over-expressing transformants. The production of native proteins was lower in these transformants compared to the controls. The over-expression of Ire1 in T. reesei had no effect on foreign protein production.
Original languageEnglish
QualificationDoctor Degree
Awarding Institution
  • University of Helsinki
Supervisors/Advisors
  • Saloheimo, Markku, Supervisor
  • Penttilä, Merja, Supervisor
Award date21 Nov 2003
Place of PublicationEspoo
Publisher
Print ISBNs951-38-6239-9
Electronic ISBNs951-38-6240-2
Publication statusPublished - 2003
MoE publication typeG5 Doctoral dissertation (article)

Fingerprint

unfolded protein response
Trichoderma reesei
fungi
Aspergillus niger
Saccharomyces cerevisiae
proteins
genes
yeasts
gene expression
protein folding
exocytosis
endocytosis
threonine
serine
molecular cloning
phosphotransferases (kinases)
organisms

Keywords

  • filamentous fungi
  • Trichoderma reesei
  • Saccharomyces cerevisiae
  • cloning
  • characterization
  • genes
  • unfolded protein response
  • gene expression
  • protein synthesis
  • protein secretion

Cite this

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title = "Functional studies of the secretory pathway of filamentous fungi: The effect of unfolded protein response on protein production: Dissertation",
abstract = "This study describes the cloning and characterisation of three genes from the filamentous fungus Trichoderma reesei. Snc1 encoding a putative v-SNARE was shown to be the functional homologue of Saccharomyces cerevisiae SNC1 and SNC2 genes that encode proteins shown to function in exo- and endocytosis. Two components of the unfolded protein response (UPR) pathway have also been identified. Ire1 encodes a serine /threonine kinase that has been shown to be involved the sensing of the protein folding status in the ER and transferring the signal to the nucleus. Ptc2 on the other hand, encodes a putative negative regulator of UPR. Both genes were isolated from T. reesei and shown to be the functional homologues of S. cerevisiae genes. The effects of UPR activation on gene expression and protein production in S. cerevisiae and two filamentous fungi, T. reesei and Aspergillus niger var. awamori are also presented. The data shows that the UPR induction, either by HacA or Ire1 overexpression in A. niger var awamori and T. reesei, respectively, induces the expression of an ER-resident foldase and an ER-resident chaperone. Moreover, the UPR induction induces the expression of genes encoding functions at different steps of the secretory pathway in both fungi. This is in correlation with results obtained in other organisms. The UPR induction was in this study shown to induce production of secreted proteins both in S. cerevisiae and A. niger var. awamori. In yeast the UPR induction by constitutive over-expression of activated yeast HAC1 and T. reesei hac1 resulted in induction of both native and foreign protein production. On the other hand, deletion of HAC1 resulted in decrease in protein production. In A. niger var. awamori only the production of foreign proteins was induced in HacA over-expressing transformants. The production of native proteins was lower in these transformants compared to the controls. The over-expression of Ire1 in T. reesei had no effect on foreign protein production.",
keywords = "filamentous fungi, Trichoderma reesei, Saccharomyces cerevisiae, cloning, characterization, genes, unfolded protein response, gene expression, protein synthesis, protein secretion",
author = "Mari Valkonen",
year = "2003",
language = "English",
isbn = "951-38-6239-9",
series = "VTT Publications",
publisher = "VTT Technical Research Centre of Finland",
number = "505",
address = "Finland",
school = "University of Helsinki",

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Functional studies of the secretory pathway of filamentous fungi : The effect of unfolded protein response on protein production: Dissertation. / Valkonen, Mari.

Espoo : VTT Technical Research Centre of Finland, 2003.

Research output: ThesisDissertationCollection of Articles

TY - THES

T1 - Functional studies of the secretory pathway of filamentous fungi

T2 - The effect of unfolded protein response on protein production: Dissertation

AU - Valkonen, Mari

PY - 2003

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N2 - This study describes the cloning and characterisation of three genes from the filamentous fungus Trichoderma reesei. Snc1 encoding a putative v-SNARE was shown to be the functional homologue of Saccharomyces cerevisiae SNC1 and SNC2 genes that encode proteins shown to function in exo- and endocytosis. Two components of the unfolded protein response (UPR) pathway have also been identified. Ire1 encodes a serine /threonine kinase that has been shown to be involved the sensing of the protein folding status in the ER and transferring the signal to the nucleus. Ptc2 on the other hand, encodes a putative negative regulator of UPR. Both genes were isolated from T. reesei and shown to be the functional homologues of S. cerevisiae genes. The effects of UPR activation on gene expression and protein production in S. cerevisiae and two filamentous fungi, T. reesei and Aspergillus niger var. awamori are also presented. The data shows that the UPR induction, either by HacA or Ire1 overexpression in A. niger var awamori and T. reesei, respectively, induces the expression of an ER-resident foldase and an ER-resident chaperone. Moreover, the UPR induction induces the expression of genes encoding functions at different steps of the secretory pathway in both fungi. This is in correlation with results obtained in other organisms. The UPR induction was in this study shown to induce production of secreted proteins both in S. cerevisiae and A. niger var. awamori. In yeast the UPR induction by constitutive over-expression of activated yeast HAC1 and T. reesei hac1 resulted in induction of both native and foreign protein production. On the other hand, deletion of HAC1 resulted in decrease in protein production. In A. niger var. awamori only the production of foreign proteins was induced in HacA over-expressing transformants. The production of native proteins was lower in these transformants compared to the controls. The over-expression of Ire1 in T. reesei had no effect on foreign protein production.

AB - This study describes the cloning and characterisation of three genes from the filamentous fungus Trichoderma reesei. Snc1 encoding a putative v-SNARE was shown to be the functional homologue of Saccharomyces cerevisiae SNC1 and SNC2 genes that encode proteins shown to function in exo- and endocytosis. Two components of the unfolded protein response (UPR) pathway have also been identified. Ire1 encodes a serine /threonine kinase that has been shown to be involved the sensing of the protein folding status in the ER and transferring the signal to the nucleus. Ptc2 on the other hand, encodes a putative negative regulator of UPR. Both genes were isolated from T. reesei and shown to be the functional homologues of S. cerevisiae genes. The effects of UPR activation on gene expression and protein production in S. cerevisiae and two filamentous fungi, T. reesei and Aspergillus niger var. awamori are also presented. The data shows that the UPR induction, either by HacA or Ire1 overexpression in A. niger var awamori and T. reesei, respectively, induces the expression of an ER-resident foldase and an ER-resident chaperone. Moreover, the UPR induction induces the expression of genes encoding functions at different steps of the secretory pathway in both fungi. This is in correlation with results obtained in other organisms. The UPR induction was in this study shown to induce production of secreted proteins both in S. cerevisiae and A. niger var. awamori. In yeast the UPR induction by constitutive over-expression of activated yeast HAC1 and T. reesei hac1 resulted in induction of both native and foreign protein production. On the other hand, deletion of HAC1 resulted in decrease in protein production. In A. niger var. awamori only the production of foreign proteins was induced in HacA over-expressing transformants. The production of native proteins was lower in these transformants compared to the controls. The over-expression of Ire1 in T. reesei had no effect on foreign protein production.

KW - filamentous fungi

KW - Trichoderma reesei

KW - Saccharomyces cerevisiae

KW - cloning

KW - characterization

KW - genes

KW - unfolded protein response

KW - gene expression

KW - protein synthesis

KW - protein secretion

M3 - Dissertation

SN - 951-38-6239-9

T3 - VTT Publications

PB - VTT Technical Research Centre of Finland

CY - Espoo

ER -