Endoplasmic reticulum stress leads to the selective transcriptional down-regulation of the glucoamylase gene in Aspergillus niger

Hashem Al-Sheikh, Adrian J. Watson, Georgina A. Lacey, Peter J. Punt, Donald A. MacKenzie, David J. Jeenes, Tiina Pakula, Merja Penttilä, Marcos J. C. Alcocer, David B. Archer

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

Abstract

We describe a new endoplasmic reticulum-associated stress response in the filamentous fungus Aspergillus niger. The inhibition of protein folding within the endoplasmic reticulum (ER) leads to cellular responses known collectively as the unfolded protein response (UPR) and we show that the selective transcriptional down-regulation of the gene encoding glucoamylase, a secreted protein, but not non-secreted proteins, is an additional consequence of ER stress. The inhibition of protein folding in the ER can be induced in a variety of ways. We have examined the effects of dithiothreitol (DTT), a reducing agent that causes the formation of unfolded proteins and have used antisense technology to lower the level of protein disulfide isomerase (PDI) in the ER of A. niger. We show that both approaches cause the down-regulation of transcription in genes encoding secreted glucoamylase and also aspergillopepsin but not genes encoding the non-secreted proteins -actin and glyceraldehyde 3' phosphate dehydrogenase. The DTT-treated fungal cells also show evidence for the induction of the UPR because expression of bipA and pdiA, encoding an ER-resident chaperone and foldase respectively, are up-regulated and splicing of hacA, the gene encoding the transcription factor responsible for induction of the UPR, occurs, allowing the production of an active protein. This response is not evident in the pdiA antisense strains, suggesting that the transcriptional down-regulation mechanism is controlled differently to the UPR. An analysis of the promoter of the glucoamylase gene using truncated glaA promoters to drive the -glucuronidase reporter gene has shown that the down regulation effect is attenuated with a promoter length of 1 kb but that the glaA promoter of 2kb exhibited the effect, suggesting that the motif(s) which mediate the response are situated within the region 1-2kb from the ATG.
Original languageEnglish
Title of host publicationPoster Abstracts
Pages196
Publication statusPublished - 2004
Event7th European Conference on Fungal Genetics - Copenhagen, Denmark
Duration: 17 Apr 200420 Apr 2004

Conference

Conference7th European Conference on Fungal Genetics
CountryDenmark
CityCopenhagen
Period17/04/0420/04/04

Fingerprint

Glucan 1,4-alpha-Glucosidase
Endoplasmic Reticulum Stress
Aspergillus niger
Unfolded Protein Response
Down-Regulation
Endoplasmic Reticulum
Genes
Dithiothreitol
Protein Folding
Proteins
Protein Disulfide-Isomerases
Protein Unfolding
Glyceraldehyde-3-Phosphate Dehydrogenases
Glucuronidase
Reducing Agents
Reporter Genes
Actins
Fungi
Transcription Factors
Technology

Cite this

Al-Sheikh, H., Watson, A. J., Lacey, G. A., Punt, P. J., MacKenzie, D. A., Jeenes, D. J., ... Archer, D. B. (2004). Endoplasmic reticulum stress leads to the selective transcriptional down-regulation of the glucoamylase gene in Aspergillus niger. In Poster Abstracts (pp. 196). [VIIIp-19]
Al-Sheikh, Hashem ; Watson, Adrian J. ; Lacey, Georgina A. ; Punt, Peter J. ; MacKenzie, Donald A. ; Jeenes, David J. ; Pakula, Tiina ; Penttilä, Merja ; Alcocer, Marcos J. C. ; Archer, David B. / Endoplasmic reticulum stress leads to the selective transcriptional down-regulation of the glucoamylase gene in Aspergillus niger. Poster Abstracts. 2004. pp. 196
@inbook{95481613678843829eda016276dc235d,
title = "Endoplasmic reticulum stress leads to the selective transcriptional down-regulation of the glucoamylase gene in Aspergillus niger",
abstract = "We describe a new endoplasmic reticulum-associated stress response in the filamentous fungus Aspergillus niger. The inhibition of protein folding within the endoplasmic reticulum (ER) leads to cellular responses known collectively as the unfolded protein response (UPR) and we show that the selective transcriptional down-regulation of the gene encoding glucoamylase, a secreted protein, but not non-secreted proteins, is an additional consequence of ER stress. The inhibition of protein folding in the ER can be induced in a variety of ways. We have examined the effects of dithiothreitol (DTT), a reducing agent that causes the formation of unfolded proteins and have used antisense technology to lower the level of protein disulfide isomerase (PDI) in the ER of A. niger. We show that both approaches cause the down-regulation of transcription in genes encoding secreted glucoamylase and also aspergillopepsin but not genes encoding the non-secreted proteins -actin and glyceraldehyde 3' phosphate dehydrogenase. The DTT-treated fungal cells also show evidence for the induction of the UPR because expression of bipA and pdiA, encoding an ER-resident chaperone and foldase respectively, are up-regulated and splicing of hacA, the gene encoding the transcription factor responsible for induction of the UPR, occurs, allowing the production of an active protein. This response is not evident in the pdiA antisense strains, suggesting that the transcriptional down-regulation mechanism is controlled differently to the UPR. An analysis of the promoter of the glucoamylase gene using truncated glaA promoters to drive the -glucuronidase reporter gene has shown that the down regulation effect is attenuated with a promoter length of 1 kb but that the glaA promoter of 2kb exhibited the effect, suggesting that the motif(s) which mediate the response are situated within the region 1-2kb from the ATG.",
author = "Hashem Al-Sheikh and Watson, {Adrian J.} and Lacey, {Georgina A.} and Punt, {Peter J.} and MacKenzie, {Donald A.} and Jeenes, {David J.} and Tiina Pakula and Merja Penttil{\"a} and Alcocer, {Marcos J. C.} and Archer, {David B.}",
year = "2004",
language = "English",
pages = "196",
booktitle = "Poster Abstracts",

}

Al-Sheikh, H, Watson, AJ, Lacey, GA, Punt, PJ, MacKenzie, DA, Jeenes, DJ, Pakula, T, Penttilä, M, Alcocer, MJC & Archer, DB 2004, Endoplasmic reticulum stress leads to the selective transcriptional down-regulation of the glucoamylase gene in Aspergillus niger. in Poster Abstracts., VIIIp-19, pp. 196, 7th European Conference on Fungal Genetics, Copenhagen, Denmark, 17/04/04.

Endoplasmic reticulum stress leads to the selective transcriptional down-regulation of the glucoamylase gene in Aspergillus niger. / Al-Sheikh, Hashem; Watson, Adrian J.; Lacey, Georgina A.; Punt, Peter J.; MacKenzie, Donald A.; Jeenes, David J.; Pakula, Tiina; Penttilä, Merja; Alcocer, Marcos J. C.; Archer, David B.

Poster Abstracts. 2004. p. 196 VIIIp-19.

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

TY - CHAP

T1 - Endoplasmic reticulum stress leads to the selective transcriptional down-regulation of the glucoamylase gene in Aspergillus niger

AU - Al-Sheikh, Hashem

AU - Watson, Adrian J.

AU - Lacey, Georgina A.

AU - Punt, Peter J.

AU - MacKenzie, Donald A.

AU - Jeenes, David J.

AU - Pakula, Tiina

AU - Penttilä, Merja

AU - Alcocer, Marcos J. C.

AU - Archer, David B.

PY - 2004

Y1 - 2004

N2 - We describe a new endoplasmic reticulum-associated stress response in the filamentous fungus Aspergillus niger. The inhibition of protein folding within the endoplasmic reticulum (ER) leads to cellular responses known collectively as the unfolded protein response (UPR) and we show that the selective transcriptional down-regulation of the gene encoding glucoamylase, a secreted protein, but not non-secreted proteins, is an additional consequence of ER stress. The inhibition of protein folding in the ER can be induced in a variety of ways. We have examined the effects of dithiothreitol (DTT), a reducing agent that causes the formation of unfolded proteins and have used antisense technology to lower the level of protein disulfide isomerase (PDI) in the ER of A. niger. We show that both approaches cause the down-regulation of transcription in genes encoding secreted glucoamylase and also aspergillopepsin but not genes encoding the non-secreted proteins -actin and glyceraldehyde 3' phosphate dehydrogenase. The DTT-treated fungal cells also show evidence for the induction of the UPR because expression of bipA and pdiA, encoding an ER-resident chaperone and foldase respectively, are up-regulated and splicing of hacA, the gene encoding the transcription factor responsible for induction of the UPR, occurs, allowing the production of an active protein. This response is not evident in the pdiA antisense strains, suggesting that the transcriptional down-regulation mechanism is controlled differently to the UPR. An analysis of the promoter of the glucoamylase gene using truncated glaA promoters to drive the -glucuronidase reporter gene has shown that the down regulation effect is attenuated with a promoter length of 1 kb but that the glaA promoter of 2kb exhibited the effect, suggesting that the motif(s) which mediate the response are situated within the region 1-2kb from the ATG.

AB - We describe a new endoplasmic reticulum-associated stress response in the filamentous fungus Aspergillus niger. The inhibition of protein folding within the endoplasmic reticulum (ER) leads to cellular responses known collectively as the unfolded protein response (UPR) and we show that the selective transcriptional down-regulation of the gene encoding glucoamylase, a secreted protein, but not non-secreted proteins, is an additional consequence of ER stress. The inhibition of protein folding in the ER can be induced in a variety of ways. We have examined the effects of dithiothreitol (DTT), a reducing agent that causes the formation of unfolded proteins and have used antisense technology to lower the level of protein disulfide isomerase (PDI) in the ER of A. niger. We show that both approaches cause the down-regulation of transcription in genes encoding secreted glucoamylase and also aspergillopepsin but not genes encoding the non-secreted proteins -actin and glyceraldehyde 3' phosphate dehydrogenase. The DTT-treated fungal cells also show evidence for the induction of the UPR because expression of bipA and pdiA, encoding an ER-resident chaperone and foldase respectively, are up-regulated and splicing of hacA, the gene encoding the transcription factor responsible for induction of the UPR, occurs, allowing the production of an active protein. This response is not evident in the pdiA antisense strains, suggesting that the transcriptional down-regulation mechanism is controlled differently to the UPR. An analysis of the promoter of the glucoamylase gene using truncated glaA promoters to drive the -glucuronidase reporter gene has shown that the down regulation effect is attenuated with a promoter length of 1 kb but that the glaA promoter of 2kb exhibited the effect, suggesting that the motif(s) which mediate the response are situated within the region 1-2kb from the ATG.

M3 - Conference abstract in proceedings

SP - 196

BT - Poster Abstracts

ER -

Al-Sheikh H, Watson AJ, Lacey GA, Punt PJ, MacKenzie DA, Jeenes DJ et al. Endoplasmic reticulum stress leads to the selective transcriptional down-regulation of the glucoamylase gene in Aspergillus niger. In Poster Abstracts. 2004. p. 196. VIIIp-19