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Eukaryotic Cell, November 2008, p. 1906-1915, Vol. 7, No. 11
1535-9778/08/$08.00+0     doi:10.1128/EC.00148-08
Copyright © 2008, American Society for Microbiology. All Rights Reserved.

C-Terminal Signals Regulate Targeting of Glycosylphosphatidylinositol-Anchored Proteins to the Cell Wall or Plasma Membrane in Candida albicans{triangledown}

Yuxin Mao,1,2 Zimei Zhang,2 Charles Gast,1 and Brian Wong1*

Division of Infectious Diseases, Oregon Health and Science University, Portland, Oregon,1 Molecular, Cellular and Developmental Biology, Yale University, New Haven, Connecticut2

Received 26 April 2008/ Accepted 15 August 2008

Fungal glycosylphosphatidylinositol (GPI)-anchored proteins localize to the plasma membrane (PM), cell wall (CW), or both. To study signals that regulate PM versus CW targeting in Candida albicans, we (i) fused the N and/or C termini of the GPI CW protein Hwp1p and the GPI PM protein Ecm331p to green fluorescent protein (GFP) and (ii) expressed and localized the resulting fusions. Forty-seven amino acids from the C terminus of Hwp1p were sufficient to target GFP to the CW, and 66 amino acids from the C terminus of Ecm331p were sufficient to target GFP to the PM. Truncation and mutagenesis studies showed that G390 was the {omega} cleavage site in Ecm331p. Domain exchange and mutagenesis studies showed that (i) the 5 amino acids immediately N-terminal to the {omega} sites (the {omega} – 5 to {omega} – 1 amino acids) played key roles in targeting to the PM or CW; (ii) KK and FE residues at positions {omega} – 1 and {omega} – 2, respectively, targeted to the PM and CW; and (iii) a loss of I at position {omega} 5 increased PM retention. Small fluorescent reporters can be used to study the peptide signals that regulate PM versus CW targeting of GPI proteins and may be useful for identifying proteins that interact with key targeting signals.


* Corresponding author. Mailing address: Division of Infectious Diseases, Oregon Health and Science University, 3181 SW Sam Jackson Park Drive (NRC-3), Portland, OR 97239. Phone: (503) 494-7735. Fax: (503) 494-4264. E-mail: wongbri{at}ohsu.edu

{triangledown} Published ahead of print on 22 August 2008.


Eukaryotic Cell, November 2008, p. 1906-1915, Vol. 7, No. 11
1535-9778/08/$08.00+0     doi:10.1128/EC.00148-08
Copyright © 2008, American Society for Microbiology. All Rights Reserved.




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