This Article Full Text (PDF) Alert me when this article is cited Alert me if a correction is posted Services Similar articles in this journal Similar articles in PubMed Alert me to new issues of the journal Download to citation manager Reprints and Permissions Copyright Information Books from ASM Press MicrobeWorld Citing Articles Citing Articles via HighWire Citing Articles via Google Scholar Google Scholar Articles by Tabuchi, M. Articles by Takegawa, K. Search for Related Content PubMed PubMed Citation Articles by Tabuchi, M. Articles by Takegawa, K.

 Previous Article  |  Next Article 

J. Bacteriol., 07 1997, 4179-4189, Vol 179, No. 13
Copyright © 1997, American Society for Microbiology

Vacuolar protein sorting in fission yeast: cloning, biosynthesis, transport, and processing of carboxypeptidase Y from Schizosaccharomyces pombe

M Tabuchi, O Iwaihara, Y Ohtani, N Ohuchi, J Sakurai, T Morita, S Iwahara and K Takegawa
Department of Bioresource Science, Faculty of Agriculture, Kagawa University, Miki-cho, Japan.

PCR was used to isolate a carboxypeptidase Y (CPY) homolog gene from the fission yeast Schizosaccharomyces pombe. The cloned S. pombe cpy1+ gene has a single open reading frame, which encodes 950 amino acids with one potential N-glycosylation site. It appears to be synthesized as an inactive pre-pro protein that likely undergoes processing following translocation into appropriate intracellular organelles. The C-terminal mature region is highly conserved in other serine carboxypeptidases. In contrast, the N-terminal pro region containing the vacuolar sorting signal in CPY from Saccharomyces cerevisiae shows fewer identical residues. The pro region contains two unusual repeating sequences; repeating sequence I consists of seven contiguous repeating segments of 13 amino acids each, and repeating sequence II consists of seven contiguous repeating segments of 9 amino acids each. Pulse-chase radiolabeling analysis revealed that Cpy1p was initially synthesized in a 110-kDa pro-precursor form and via the 51-kDa single-polypeptide- chain intermediate form which has had its pro segment removed is finally converted to a heterodimer, the mature form, which is detected as a 32-kDa protein on sodium dodecyl sulfate-polyacrylamide gel electrophoresis under reducing conditions. Like S. cerevisiae CPY, S. pombe Cpy1p does not require the N-linked oligosaccharide moiety for vacuolar delivery. To investigate the vacuolar sorting signal of S. pombe Cpy1p, we have constructed cpy1+-SUC2 gene fusions that direct the synthesis of hybrid proteins consisting of N-terminal segments of various lengths of S. pombe Cpy1p fused to the secreted enzyme S. cerevisiae invertase. The N-terminal 478 amino acids of Cpy1 are sufficient to direct delivery of a Cpy1-Inv hybrid protein to the vacuole. These results showed that the pro peptide of Cpy1 contains the putative vacuolar sorting signal.

This article has been cited by other articles:

• Codlin, S., Mole, S. E. (2009). S. pombe btn1, the orthologue of the Batten disease gene CLN3, is required for vacuole protein sorting of Cpy1p and Golgi exit of Vps10p. J. Cell Sci. 122: 1163-1173 [Abstract] [Full Text]  
• Iwaki, T., Iefuji, H., Hiraga, Y., Hosomi, A., Morita, T., Giga-Hama, Y., Takegawa, K. (2008). Multiple functions of ergosterol in the fission yeast Schizosaccharomyces pombe. Microbiology 154: 830-841 [Abstract] [Full Text]  
• Iwaki, T., Onishi, M., Ikeuchi, M., Kita, A., Sugiura, R., Giga-Hama, Y., Fukui, Y., Takegawa, K. (2007). Essential roles of class E Vps proteins for sorting into multivesicular bodies in Schizosaccharomyces pombe. Microbiology 153: 2753-2764 [Abstract] [Full Text]  
• Miyatake, M., Kuno, T., Kita, A., Katsura, K., Takegawa, K., Uno, S., Nabata, T., Sugiura, R. (2007). Valproic Acid Affects Membrane Trafficking and Cell-Wall Integrity in Fission Yeast. Genetics 175: 1695-1705 [Abstract] [Full Text]  
• Iwaki, T., Hosomi, A., Tokudomi, S., Kusunoki, Y., Fujita, Y., Giga-Hama, Y., Tanaka, N., Takegawa, K. (2006). Vacuolar protein sorting receptor in Schizosaccharomyces pombe.. Microbiology 152: 1523-1532 [Abstract] [Full Text]  
• Koga, T., Onishi, M., Nakamura, Y., Hirata, A., Nakamura, T., Shimoda, C., Iwaki, T., Takegawa, K., Fukui, Y. (2004). Sorting nexin homologues are targets of phosphatidylinositol 3-phosphate in sporulation of Schizosaccharomyces pombe. GENES CELLS 9: 561-574 [Abstract] [Full Text]  
• Maeda, T., Sugiura, R., Kita, A., Saito, M., Deng, L., He, Y., Lu, Y., Fujita, Y., Takegawa, K., Shuntoh, H., Kuno, T. (2004). Pmr1, a P-type ATPase, and Pdt1, an Nramp homologue, cooperatively regulate cell morphogenesis in fission yeast: The importance of Mn2+ homeostasis. GENES CELLS 9: 71-82 [Abstract] [Full Text]  
• Skrzypek, E., Myers-Morales, T., Whiteheart, S. W., Straley, S. C. (2003). Application of a Saccharomyces cerevisiae Model To Study Requirements for Trafficking of Yersinia pestis YopM in Eucaryotic Cells. Infect. Immun. 71: 937-947 [Abstract] [Full Text]  
• Cheng, H., Sugiura, R., Wu, W., Fujita, M., Lu, Y., Sio, S. O., Kawai, R., Takegawa, K., Shuntoh, H., Kuno, T. (2002). Role of the Rab GTP-Binding Protein Ypt3 in the Fission Yeast Exocytic Pathway and Its Connection to Calcineurin Function. Mol. Biol. Cell 13: 2963-2976 [Abstract] [Full Text]  
• D'Alessio, C., Fernandez, F., Trombetta, E. S., Parodi, A. J. (1999). Genetic Evidence for the Heterodimeric Structure of Glucosidase II. THE EFFECT OF DISRUPTING THE SUBUNIT-ENCODING GENES ON GLYCOPROTEIN FOLDING. J. Biol. Chem. 274: 25899-25905 [Abstract] [Full Text]  
• Fanchiotti, S., Fernandez, F., D'Alessio, C., Parodi, A. J. (1998). The UDP-Glc:Glycoprotein Glucosyltransferase Is Essential for Schizosaccharomyces pombe Viability under Conditions of Extreme Endoplasmic Reticulum Stress. JCB 143: 625-635 [Abstract] [Full Text]  
• Ingavale, S. S., Kaur, R., Aggarwal, P., Bachhawat, A. K. (1998). A Minisatellite Sequence within the Propeptide Region of the Vacuolar Carboxypeptidase Y Gene of Schizosaccharomyces pombe. J. Bacteriol. 180: 3727-3729 [Abstract] [Full Text]