TY - JOUR
T1 - Comparative topology studies in Saccharomyces cerevisiae and in Escherichia coli
T2 - The N-terminal half of the yeast ABC protein Ste6
AU - Geller, Dorit
AU - Taglicht, Daniel
AU - Edgar, Rotem
AU - Tam, Amy
AU - Pines, Ophry
AU - Michaelis, Susan
AU - Bibi, Eitan
PY - 1996
Y1 - 1996
N2 - 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.
AB - 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.
UR - http://www.scopus.com/inward/record.url?scp=0030011712&partnerID=8YFLogxK
U2 - 10.1074/jbc.271.23.13746
DO - 10.1074/jbc.271.23.13746
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C2 - 8662764
AN - SCOPUS:0030011712
SN - 0021-9258
VL - 271
SP - 13746
EP - 13753
JO - Journal of Biological Chemistry
JF - Journal of Biological Chemistry
IS - 23
ER -