Mutational analysis of the yeast a-factor transporter STE6, a member of the ATP binding cassette (ABC) protein superfamily

Carol Berkower, Susan Doris Michaelis

Research output: Contribution to journalArticle

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 languageEnglish (US)
Pages (from-to)3777-3785
Number of pages9
JournalThe EMBO journal
Volume10
Issue number12
StatePublished - 1991

Fingerprint

Yeast
Carrier Proteins
Adenosine Triphosphate
Yeasts
Cystic Fibrosis
Nucleotides
P-Glycoproteins
Mutation
Proteins
Molecules
Genes
Membranes
Peptides

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 journalArticle

@article{00899f9e92d943ec9a5eb8527eccad48,
title = "Mutational analysis of the yeast a-factor transporter STE6, a member of the ATP binding cassette (ABC) protein superfamily",
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.",
keywords = "A-factor mating pheromone transporter, ATP binding protein, CFTR, MDR, Yeast STE6",
author = "Carol Berkower and Michaelis, {Susan Doris}",
year = "1991",
language = "English (US)",
volume = "10",
pages = "3777--3785",
journal = "EMBO Journal",
issn = "0261-4189",
publisher = "Nature Publishing Group",
number = "12",

}

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

UR - http://www.scopus.com/inward/record.url?scp=0026094617&partnerID=8YFLogxK

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 -