A Regulatory Domain in the C-terminal Extension of the Yeast Glycerol Channel Fps1p
Artikel i vetenskaplig tidskrift, 2004

The Saccharomyces cerevisiae gene FPS1 encodes an aquaglyceroporin of the major intrinsic protein (MIP) family. The main function of Fps1p seems to be the efflux of glycerol in the adaptation of the yeast cell to lower external osmolarity. Fps1p is an atypical member of the family, because the protein is much larger (669 amino acids) than most MIPs due to long hydrophilic extensions in both termini. We have shown previously that a short domain in the N-terminal extension of the protein is required for restricting glycerol transport through the channel (Tamás, M. J., Karlgren, S., Bill, R. M., Hedfalk, K., Allegri, L., Ferreira, M., Thevelein, J. M., Rydström, J., Mullins, J. G. L., and Hohmann, S. (2003) J. Biol. Chem. 278, 6337–6345). Deletion of the N-terminal domain results in an unregulated channel, loss of glycerol, and osmosensitivity. In this work we have investigated the role of the Fps1p C terminus (139 amino acids). A set of eight truncations has been constructed and tested in vivo in a yeast fps1 strain. We have performed growth tests, membrane localization following cell fractionation, and glycerol accumulation measurements as well as an investigation of the osmotic stress response. Our results show that the C-terminal extension is also involved in restricting transport through Fps1p. We have identified a sequence of 12 amino acids, residues 535–546, close to the sixth transmembrane domain. This element seems to be important for controlling Fps1p function. Similar to the N-terminal domain, the C-terminal domain is amphiphilic and has a potential to dip into the membrane

Författare

Kristina Hedfalk

Göteborgs universitet

R Bill

Johathan G.L. Mullins

Sara Karlgren

Göteborgs universitet

Caroline Filipsson

Göteborgs universitet

Johanna Bergström

Markus J. Tamás

Göteborgs universitet

Jan Rydström

Göteborgs universitet

Stefan Hohmann

Göteborgs universitet

Journal of biological chemistry

Vol. 279 14954-14960

Ämneskategorier

Biologiska vetenskaper

DOI

10.1074/jbc.M313126200