Abstract
STE6, the yeast a-factor transporter, is a member of the ATP binding cassette protein superfamily, which also includes the mammalian multidrug resistance protein and the cystic fibrosis gene product. These proteins contain two homologous halves, each with six membrane spanning segments and a predicted ATP nucleotide binding domain. To assess the importance of the two halves of STE6, and to examine the functional significance of residues conserved among members of the ATP binding cassette superfamily, we introduced mutations into the nucleotide binding domains of STE6. Our analysis demonstrates that both halves of STE6 are critical for function and that some, but not all, mutations analogous to those known to result in cystic fibrosis impair STE6 activity. To examine further the functional contribution of each half of the STE6 protein, we severed the STE6 coding sequence and expressed the two halves of the transporter as separate polypeptides. Whereas 'half-molecules' are unable to provide transport function individually, co-expression of both half-molecules in the same cell leads to functional reconstitution of STE6-mediated a-factor transport.
| Original language | English (US) |
|---|---|
| Pages (from-to) | 3777-3785 |
| Number of pages | 9 |
| Journal | The EMBO journal |
| Volume | 10 |
| Issue number | 12 |
| State | Published - 1991 |
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Keywords
- A-factor mating pheromone transporter
- ATP binding protein
- CFTR
- MDR
- Yeast STE6
ASJC Scopus subject areas
- Cell Biology
- Genetics
Cite this
Mutational analysis of the yeast a-factor transporter STE6, a member of the ATP binding cassette (ABC) protein superfamily. / Berkower, Carol; Michaelis, Susan Doris.
In: The EMBO journal, Vol. 10, No. 12, 1991, p. 3777-3785.Research output: Contribution to journal › Article
}
TY - JOUR
T1 - Mutational analysis of the yeast a-factor transporter STE6, a member of the ATP binding cassette (ABC) protein superfamily
AU - Berkower, Carol
AU - Michaelis, Susan Doris
PY - 1991
Y1 - 1991
N2 - STE6, the yeast a-factor transporter, is a member of the ATP binding cassette protein superfamily, which also includes the mammalian multidrug resistance protein and the cystic fibrosis gene product. These proteins contain two homologous halves, each with six membrane spanning segments and a predicted ATP nucleotide binding domain. To assess the importance of the two halves of STE6, and to examine the functional significance of residues conserved among members of the ATP binding cassette superfamily, we introduced mutations into the nucleotide binding domains of STE6. Our analysis demonstrates that both halves of STE6 are critical for function and that some, but not all, mutations analogous to those known to result in cystic fibrosis impair STE6 activity. To examine further the functional contribution of each half of the STE6 protein, we severed the STE6 coding sequence and expressed the two halves of the transporter as separate polypeptides. Whereas 'half-molecules' are unable to provide transport function individually, co-expression of both half-molecules in the same cell leads to functional reconstitution of STE6-mediated a-factor transport.
AB - STE6, the yeast a-factor transporter, is a member of the ATP binding cassette protein superfamily, which also includes the mammalian multidrug resistance protein and the cystic fibrosis gene product. These proteins contain two homologous halves, each with six membrane spanning segments and a predicted ATP nucleotide binding domain. To assess the importance of the two halves of STE6, and to examine the functional significance of residues conserved among members of the ATP binding cassette superfamily, we introduced mutations into the nucleotide binding domains of STE6. Our analysis demonstrates that both halves of STE6 are critical for function and that some, but not all, mutations analogous to those known to result in cystic fibrosis impair STE6 activity. To examine further the functional contribution of each half of the STE6 protein, we severed the STE6 coding sequence and expressed the two halves of the transporter as separate polypeptides. Whereas 'half-molecules' are unable to provide transport function individually, co-expression of both half-molecules in the same cell leads to functional reconstitution of STE6-mediated a-factor transport.
KW - A-factor mating pheromone transporter
KW - ATP binding protein
KW - CFTR
KW - MDR
KW - Yeast STE6
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UR - http://www.scopus.com/inward/citedby.url?scp=0026094617&partnerID=8YFLogxK
M3 - Article
C2 - 1935899
AN - SCOPUS:0026094617
VL - 10
SP - 3777
EP - 3785
JO - EMBO Journal
JF - EMBO Journal
SN - 0261-4189
IS - 12
ER -