Identification of a cytoplasm to vacuole targeting determinant in aminopeptidase I

doi:10.1083/jcb.132.6.999

PeproTech: Free Shipping at www.peprotech.com

This Article

	Full Text (PDF, 1765K)
	Alert me when this article is cited
	Citation Map

Services

	Email this article
	Similar articles in this journal
	Similar articles in PubMed
	Alert me to new content in the JCB
	Download to citation manager

Citing Articles

	Citing Articles via HighWire
	Citing Articles via CrossRef
	Citing Articles via Google Scholar

Google Scholar

	Articles by Oda, M. N.
	Articles by Klionsky, D. J.
	Search for Related Content

PubMed

	PubMed Citation
	Articles by Oda, M. N.
	Articles by Klionsky, D. J.


Social Bookmarking

	What's this?

ARTICLES

Identification of a cytoplasm to vacuole targeting determinant in aminopeptidase I

MN Oda, SV Scott, A Hefner-Gravink, AD Caffarelli and DJ Klionsky
Section of Microbiology, University of California, Davis 95616, USA.

Aminopeptidase I (API) is a soluble leucine aminopeptidase resident in the yeast vacuole (Frey, J., and K.H. Rohm. 1978. Biochim. Biophys. Acta. 527:31-41). The precursor form of API contains an amino-terminal 45-amino acid propeptide, which is removed by proteinase B (PrB) upon entry into the vacuole. The propeptide of API lacks a consensus signal sequence and it has been demonstrated that vacuolar localization of API is independent of the secretory pathway (Klionsky, D.J., R. Cueva, and D.S. Yaver. 1992. J. Cell Biol. 119:287-299). The predicted secondary structure for the API propeptide is composed of an amphipathic alpha- helix followed by a beta-turn and another alpha-helix, forming a helix- turn-helix structure. With the use of mutational analysis, we determined that the API propeptide is essential for proper transport into the vacuole. Deletion of the entire propeptide from the API molecule resulted in accumulation of a mature-sized protein in the cytosol. A more detailed examination using random mutagenesis and a series of smaller deletions throughout the propeptide revealed that API localization is severely affected by alterations within the predicted first alpha-helix. In vitro studies indicate that mutations in this predicted helix prevent productive binding interactions from taking place. In contrast, vacuolar import is relatively insensitive to alterations in the second predicted helix of the propeptide. Examination of API folding revealed that mutations that affect entry into the vacuole did not affect the structure of API. These data indicate that the API propeptide serves as a vacuolar targeting determinant at a critical step along the cytoplasm to vacuole targeting pathway.
Add to CiteULike CiteULike Add to Complore Complore Add to Connotea Connotea Add to Del.icio.us Del.icio.us Add to Digg Digg Facebook Add to Reddit Reddit Add to Technorati Technorati Twitter What's this?

This article has been cited by other articles:

McIntosh, M. T., Vaid, A., Hosgood, H. D., Vijay, J., Bhattacharya, A., Sahani, M. H., Baevova, P., Joiner, K. A., Sharma, P. (2007). Traffic to the Malaria Parasite Food Vacuole: A NOVEL PATHWAY INVOLVING A PHOSPHATIDYLINOSITOL 3-PHOSPHATE-BINDING PROTEIN. J. Biol. Chem. 282: 11499-11508 [Abstract] [Full Text]
Palmer, G. E., Kelly, M. N., Sturtevant, J. E. (2007). Autophagy in the pathogen Candida albicans. Microbiology 153: 51-58 [Abstract] [Full Text]
He, C., Song, H., Yorimitsu, T., Monastyrska, I., Yen, W.-L., Legakis, J. E., Klionsky, D. J. (2006). Recruitment of Atg9 to the preautophagosomal structure by Atg11 is essential for selective autophagy in budding yeast. JCB 175: 925-935 [Abstract] [Full Text]
Reggiori, F., Monastyrska, I., Shintani, T., Klionsky, D. J. (2005). The Actin Cytoskeleton Is Required for Selective Types of Autophagy, but Not Nonspecific Autophagy, in the Yeast Saccharomyces cerevisiae. Mol. Biol. Cell 16: 5843-5856 [Abstract] [Full Text]
Yorimitsu, T., Klionsky, D. J. (2005). Atg11 Links Cargo to the Vesicle-forming Machinery in the Cytoplasm to Vacuole Targeting Pathway. Mol. Biol. Cell 16: 1593-1605 [Abstract] [Full Text]
Guan, J., Stromhaug, P. E., George, M. D., Habibzadegah-Tari, P., Bevan, A., Dunn, W. A. Jr., Klionsky, D. J. (2001). Cvt18/Gsa12 Is Required for Cytoplasm-to-Vacuole Transport, Pexophagy, and Autophagy in Saccharomyces cerevisiae and Pichia pastoris. Mol. Biol. Cell 12: 3821-3838 [Abstract] [Full Text]
Kim, J., Kamada, Y., Stromhaug, P. E., Guan, J., Hefner-Gravink, A., Baba, M., Scott, S. V., Ohsumi, Y., Dunn, W. A. Jr., Klionsky, D. J. (2001). Cvt9/Gsa9 Functions in Sequestering Selective Cytosolic Cargo Destined for the Vacuole. JCB 153: 381-396 [Abstract] [Full Text]
Kim, J., Dalton, V. M., Eggerton, K. P., Scott, S. V., Klionsky, D. J. (1999). Apg7p/Cvt2p Is Required for the Cytoplasm-to-Vacuole Targeting, Macroautophagy, and Peroxisome Degradation Pathways. Mol. Biol. Cell 10: 1337-1351 [Abstract] [Full Text]
Hill, K., Hutchings, N., Russell, D., Donelson, J. (1999). A novel protein targeting domain directs proteins to the anterior cytoplasmic face of the flagellar pocket in African trypanosomes. J. Cell Sci. 112: 3091-3101 [Abstract]
Klionsky, D. J. (1998). Nonclassical Protein Sorting to the Yeast Vacuole. J. Biol. Chem. 273: 10807-10810 [Full Text]
Bryant, N. J., Stevens, T. H. (1998). Vacuole Biogenesis in Saccharomyces cerevisiae: Protein Transport Pathways to the Yeast Vacuole. Microbiol. Mol. Biol. Rev. 62: 230-247 [Abstract] [Full Text]
Baba, M., Osumi, M., Scott, S. V., Klionsky, D. J., Ohsumi, Y. (1997). Two Distinct Pathways for Targeting Proteins from the Cytoplasm to the Vacuole/Lysosome. JCB 139: 1687-1695 [Abstract] [Full Text]
Scott, S. V., Baba, M., Ohsumi, Y., Klionsky, D. J. (1997). Aminopeptidase I Is Targeted to the Vacuole by a Nonclassical Vesicular Mechanism. JCB 138: 37-44 [Abstract] [Full Text]
Kim, J., Scott, S. V., Oda, M. N., Klionsky, D. J. (1997). Transport of a Large Oligomeric Protein by the Cytoplasm to Vacuole Protein Targeting Pathway. JCB 137: 609-618 [Abstract] [Full Text]
Harding, T. M., Hefner-Gravink, A., Thumm, M., Klionsky, D. J. (1996). Genetic and Phenotypic Overlap between Autophagy and the Cytoplasm to Vacuole Protein Targeting Pathway. J. Biol. Chem. 271: 17621-17624 [Abstract] [Full Text]
Scott, S. V., Nice, D. C. III, Nau, J. J., Weisman, L. S., Kamada, Y., Keizer-Gunnink, I., Funakoshi, T., Veenhuis, M., Ohsumi, Y., Klionsky, D. J. (2000). Apg13p and Vac8p Are Part of a Complex of Phosphoproteins That Are Required for Cytoplasm to Vacuole Targeting. J. Biol. Chem. 275: 25840-25849 [Abstract] [Full Text]
Silles, E., Mazon, M. J., Gevaert, K., Goethals, M., Vandekerckhove, J., Leber, R., Sandoval, I. V. (2000). Targeting of Aminopeptidase I to the Yeast Vacuole Is Mediated by Ssa1p, a Cytosolic Member of the 70-kDa Stress Protein Family. J. Biol. Chem. 275: 34054-34059 [Abstract] [Full Text]
Andrei-Selmer, C., Knuppel, A., Satyanarayana, C., Heese, C., Schu, P. V. (2001). A New Class of Mutants Deficient in Dodecamerization of Aminopeptidase 1 and Vacuolar Transport. J. Biol. Chem. 276: 11606-11614 [Abstract] [Full Text]
Leber, R., Silles, E., Sandoval, I. V., Mazon, M. J. (2001). Yol082p, a Novel CVT Protein Involved in the Selective Targeting of Aminopeptidase I to the Yeast Vacuole. J. Biol. Chem. 276: 29210-29217 [Abstract] [Full Text]