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Analysis of the hypoxia-induced ADH2-promoter of the respiratory yeast Pichia stipitis reveals a new mechanism for sensing of oxygen limitation in yeast

Passoth, Volkmar; Cohn, Marita LU ; Schäfer, B.; Hahn-Hägerdal, Bärbel LU and Klinner, U. (2003) In Yeast 20(1). p.39-51
Abstract
We introduced a reporter gene system into Pichia stipitis using the gene for the artificial green fluorescent protein (GFP), variant yEGFP. This system was used to analyse hypoxia-dependent PsADH2 regulation. Reporter gene activity was only found under oxygen limitation on a fermentable carbon source. The promoter was not induced by oxygen limitation in the Crabtree-positive yeast Saccharomyces cerevisiae. Promoter deletions revealed that a region of 15 bp contained the essential site for hypoxic induction. This motif was different from the known hypoxia response elements of S. cerevisiae but showed some similarity to the mammalian HIF-1 binding site. Electrophoretic mobility shift assays demonstrated specific protein binding to this... (More)
We introduced a reporter gene system into Pichia stipitis using the gene for the artificial green fluorescent protein (GFP), variant yEGFP. This system was used to analyse hypoxia-dependent PsADH2 regulation. Reporter gene activity was only found under oxygen limitation on a fermentable carbon source. The promoter was not induced by oxygen limitation in the Crabtree-positive yeast Saccharomyces cerevisiae. Promoter deletions revealed that a region of 15 bp contained the essential site for hypoxic induction. This motif was different from the known hypoxia response elements of S. cerevisiae but showed some similarity to the mammalian HIF-1 binding site. Electrophoretic mobility shift assays demonstrated specific protein binding to this region under oxygen limitation. Similar to the S. cerevisiae heme sensor system, the promoter was induced by Co2+. Cyanide was not able to mimic the effect of oxygen limitation. The activation mechanism of PsADH2 also, in this respect, has similarities to the mammalian HIF-1 system, which is inducible by Co2+ but not by cyanide. Thus, the very first promoter analysis in P. stipitis revealed a hitherto unknown mechanism of oxygen sensing in yeast. (Less)
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author
organization
publishing date
type
Contribution to journal
publication status
published
subject
keywords
hypoxia, green fluorescent, alcohol dehydrogenase, promoter analysis, protein, oxygen limitation
in
Yeast
volume
20
issue
1
pages
39 - 51
publisher
John Wiley & Sons
external identifiers
  • scopus:0037438565
  • pmid:12489125
  • wos:000180219900005
ISSN
1097-0061
DOI
10.1002/yea.933
language
English
LU publication?
yes
id
b2b8a502-4594-4573-b6cd-7e7f2bd366eb (old id 4754507)
date added to LUP
2014-11-06 11:24:10
date last changed
2017-01-01 06:53:49
@article{b2b8a502-4594-4573-b6cd-7e7f2bd366eb,
  abstract     = {We introduced a reporter gene system into Pichia stipitis using the gene for the artificial green fluorescent protein (GFP), variant yEGFP. This system was used to analyse hypoxia-dependent PsADH2 regulation. Reporter gene activity was only found under oxygen limitation on a fermentable carbon source. The promoter was not induced by oxygen limitation in the Crabtree-positive yeast Saccharomyces cerevisiae. Promoter deletions revealed that a region of 15 bp contained the essential site for hypoxic induction. This motif was different from the known hypoxia response elements of S. cerevisiae but showed some similarity to the mammalian HIF-1 binding site. Electrophoretic mobility shift assays demonstrated specific protein binding to this region under oxygen limitation. Similar to the S. cerevisiae heme sensor system, the promoter was induced by Co2+. Cyanide was not able to mimic the effect of oxygen limitation. The activation mechanism of PsADH2 also, in this respect, has similarities to the mammalian HIF-1 system, which is inducible by Co2+ but not by cyanide. Thus, the very first promoter analysis in P. stipitis revealed a hitherto unknown mechanism of oxygen sensing in yeast.},
  author       = {Passoth, Volkmar and Cohn, Marita and Schäfer, B. and Hahn-Hägerdal, Bärbel and Klinner, U.},
  issn         = {1097-0061},
  keyword      = {hypoxia,green fluorescent,alcohol dehydrogenase,promoter analysis,protein,oxygen limitation},
  language     = {eng},
  number       = {1},
  pages        = {39--51},
  publisher    = {John Wiley & Sons},
  series       = {Yeast},
  title        = {Analysis of the hypoxia-induced ADH2-promoter of the respiratory yeast Pichia stipitis reveals a new mechanism for sensing of oxygen limitation in yeast},
  url          = {http://dx.doi.org/10.1002/yea.933},
  volume       = {20},
  year         = {2003},
}