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Journal of Bacteriology, July 1999, p. 3886-3889, Vol. 181, No. 13
0021-9193/99/$04.00+0
Copyright © 1999, American Society for Microbiology. All rights reserved.

Amino Acid Residues in the omega -Minus Region Participate in Cellular Localization of Yeast Glycosylphosphatidylinositol-Attached Proteins

Kenji Hamada, Hiromichi Terashima, Mikio Arisawa, Nami Yabuki, and Kunio Kitada*

Department of Mycology, Nippon Roche Research Center, Kamakura, Kanagawa 247-8530, Japan

Received 10 March 1999/Accepted 29 April 1999

The final destination of glycosylphosphatidylinositol (GPI)-attached proteins in Saccharomyces cerevisiae is the plasma membrane or the cell wall. Two kinds of signals have been proposed for their cellular localization: (i) the specific amino acid residues V, I, or L at the site 4 or 5 amino acids upstream of the GPI attachment site (the omega  site) and Y or N at the site 2 amino acids upstream of the omega  site for cell wall localization and (ii) dibasic residues in the region upstream of the omega  site (the omega -minus region) for plasma membrane localization. The relationships between these amino acid residues and efficiencies of cell wall incorporation were examined by constructing fusion reporter proteins from open reading frames encoding putative GPI-attached proteins. The levels of incorporation were high in the constructs containing the specific amino acid residues and quite low in those containing two basic amino acid residues in the omega -minus region. With constructs that contained neither specific residues nor two basic residues, levels of incorporation were moderate. These correlations clearly suggest that GPI-attached proteins have two different signals which act positively or negatively in cell wall incorporation for their cellular localization.

* Corresponding author. Mailing address: Department of Mycology, Nippon Roche Research Center, 200-Kajiwara, Kamakura, Kanagawa 247-8530, Japan. Phone: 81-467-45-6752. Fax: 81-467-46-5320. E-mail: kunio.kitada{at}roche.com.

Journal of Bacteriology, July 1999, p. 3886-3889, Vol. 181, No. 13
0021-9193/99/$04.00+0
Copyright © 1999, American Society for Microbiology. All rights reserved.


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