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Properties of the cysteine-less Pho84 phosphate transporter of Saccharomyces cerevisiae

Berhe, A; Zvyagilskaya, Renata; Lagerstedt, J O LU and Pratt, James R (2001) In Biochemical and biophysical research communications 287(4). p.42-837
Abstract

The derepressible Pho84 high-affinity phosphate permease of Saccharomyces cerevisiae, encoded by the PHO84 gene belongs to a family of phosphate:proton symporters (PHS). The protein contains 12 native cysteine residues of which five are predicted to be located in putative transmembrane regions III, VI, VIII, IX, and X, and the remaining seven in the hydrophilic domains of the protein. Here we report on the construction of a Pho84 transporter devoid of cysteine residues (C-less) in which all 12 native residues were replaced with serines using PCR mutagenesis and the functional consequences of this. Our results clearly demonstrate that the C-less Pho84 variant is able to support growth of yeast cells to the same extent as the wild-type... (More)

The derepressible Pho84 high-affinity phosphate permease of Saccharomyces cerevisiae, encoded by the PHO84 gene belongs to a family of phosphate:proton symporters (PHS). The protein contains 12 native cysteine residues of which five are predicted to be located in putative transmembrane regions III, VI, VIII, IX, and X, and the remaining seven in the hydrophilic domains of the protein. Here we report on the construction of a Pho84 transporter devoid of cysteine residues (C-less) in which all 12 native residues were replaced with serines using PCR mutagenesis and the functional consequences of this. Our results clearly demonstrate that the C-less Pho84 variant is able to support growth of yeast cells to the same extent as the wild-type Pho84 and is stably expressed under derepressible conditions and is fully active in proton-coupled phosphate transport across the yeast plasma membrane.

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author
publishing date
type
Contribution to journal
publication status
published
keywords
Cysteine, Fungal Proteins, Genes, myc, Hydrogen-Ion Concentration, Membrane Transport Proteins, Mutagenesis, Site-Directed, Phosphates, Protein Structure, Secondary, Proton-Phosphate Symporters, Recombinant Fusion Proteins, Saccharomyces cerevisiae, Saccharomyces cerevisiae Proteins, Journal Article, Research Support, Non-U.S. Gov't
in
Biochemical and biophysical research communications
volume
287
issue
4
pages
42 - 837
publisher
Elsevier
external identifiers
  • scopus:0035812688
ISSN
0006-291X
DOI
10.1006/bbrc.2001.5664
language
English
LU publication?
no
id
62b70b91-5bc4-48f6-8dbe-1a556b48a6a7
date added to LUP
2017-10-19 20:13:23
date last changed
2018-05-29 12:06:20
@article{62b70b91-5bc4-48f6-8dbe-1a556b48a6a7,
  abstract     = {<p>The derepressible Pho84 high-affinity phosphate permease of Saccharomyces cerevisiae, encoded by the PHO84 gene belongs to a family of phosphate:proton symporters (PHS). The protein contains 12 native cysteine residues of which five are predicted to be located in putative transmembrane regions III, VI, VIII, IX, and X, and the remaining seven in the hydrophilic domains of the protein. Here we report on the construction of a Pho84 transporter devoid of cysteine residues (C-less) in which all 12 native residues were replaced with serines using PCR mutagenesis and the functional consequences of this. Our results clearly demonstrate that the C-less Pho84 variant is able to support growth of yeast cells to the same extent as the wild-type Pho84 and is stably expressed under derepressible conditions and is fully active in proton-coupled phosphate transport across the yeast plasma membrane.</p>},
  author       = {Berhe, A and Zvyagilskaya, Renata and Lagerstedt, J O and Pratt, James R},
  issn         = {0006-291X},
  keyword      = {Cysteine,Fungal Proteins,Genes, myc,Hydrogen-Ion Concentration,Membrane Transport Proteins,Mutagenesis, Site-Directed,Phosphates,Protein Structure, Secondary,Proton-Phosphate Symporters,Recombinant Fusion Proteins,Saccharomyces cerevisiae,Saccharomyces cerevisiae Proteins,Journal Article,Research Support, Non-U.S. Gov't},
  language     = {eng},
  month        = {10},
  number       = {4},
  pages        = {42--837},
  publisher    = {Elsevier},
  series       = {Biochemical and biophysical research communications},
  title        = {Properties of the cysteine-less Pho84 phosphate transporter of Saccharomyces cerevisiae},
  url          = {http://dx.doi.org/10.1006/bbrc.2001.5664},
  volume       = {287},
  year         = {2001},
}