Comparative topology studies in Saccharomyces cerevisiae and in Escherichia coli: The N-terminal half of the yeast ABC protein Ste6

Dorit Geller, Daniel Taglicht, Rotem Edgar, Amy Tam, Ophry Pines, Susan Michaelis, Eitan Bibi

Research output: Contribution to journalArticlepeer-review

Abstract

Gene fusions have provided a strategy for determining the topology of polytopic membrane proteins in Escherichia coli. To evaluate whether this highly effective approach is applicable to heterologously expressed eukaryotic integral membrane proteins, we have carried out a comparative topological study of the eukaryotic membrane protein Ste6 both in bacteria and in yeast. Ste6, is an ATP binding cassette (ABC) protein, essential for export of the a-factor mating pheromone in Saccharomyces cerevisiae. The topogenic reporters, invertase in S. cerevisiae and alkaline phosphatase in E. coli, were fused to Ste6 at identical sites and the fusions were expressed in yeast and bacteria, respectively. The results obtained in both systems are similar, although more definitive in E. coli, and support the predicted six-transmembrane spans organization of the N-terminal half of Ste6. Thus, the topological determinants for membrane insertion of polytopic proteins in prokaryotic and in eukaryotic systems appear to be highly similar. In this study we also demonstrate that Ste6 does not contain a cleaved signal sequence.

Original languageEnglish (US)
Pages (from-to)13746-13753
Number of pages8
JournalJournal of Biological Chemistry
Volume271
Issue number23
DOIs
StatePublished - 1996

ASJC Scopus subject areas

  • Biochemistry
  • Molecular Biology
  • Cell Biology

Fingerprint

Dive into the research topics of 'Comparative topology studies in Saccharomyces cerevisiae and in Escherichia coli: The N-terminal half of the yeast ABC protein Ste6'. Together they form a unique fingerprint.

Cite this