Genetics, Vol 143, 1555-1566, Copyright © 1996

INVESTIGATIONS

Isolation of Degradation-Deficient Mutants Defective in the Targeting of Fructose-1,6-bisphosphatase into the Vacuole for Degradation in Saccharomyces cerevisiae

M. Hoffman and H. L. Chiang
Department of Cell Biology, Harvard Medical School, Boston, Massachusetts 02115

The key regulatory enzyme in the gluconeogenesis pathway, fructose-1,6-bisphosphatase (FBPase), is induced when Saccharomyces cerevisiae are grown in medium containing a poor carbon source. FBPase is targeted to the yeast vacuole for degradation when glucose-starved cells are replenished with fresh glucose. To identify genes involved in the FBPase degradation pathway, mutants that failed to degrade FBPase in response to glucose were isolated using a colony-blotting procedure. These vacuolar import and degradation-deficient (vid) mutants were placed into 20 complementation groups. They are distinct from the known sec, vps or pep mutants affecting protein secretion, vacuolar sorting and vacuolar proteolysis in that they sort CpY correctly and regulate osmotic pressure normally. Despite the presence of FBPase antigen in these mutants, FBPase is completely inactivated in all vid mutants, indicating that the c-AMP-dependent signal transduction pathway and inactivation must function properly in vid mutants. vid mutants block FBPase degradation by accumulating FBPase in the cytosol and also in small vesicles in the cytoplasm. FBPase may be targeted to small vesicles before uptake by the vacuole.

This article has been cited by other articles:

				C. R. Brown, A. B. Wolfe, D. Cui, and H.-L. Chiang The Vacuolar Import and Degradation Pathway Merges with the Endocytic Pathway to Deliver Fructose-1,6-bisphosphatase to the Vacuole for Degradation J. Biol. Chem., September 19, 2008; 283(38): 26116 - 26127. [Abstract] [Full Text] [PDF]

				O. Santt, T. Pfirrmann, B. Braun, J. Juretschke, P. Kimmig, H. Scheel, K. Hofmann, M. Thumm, and D. H. Wolf The Yeast GID Complex, a Novel Ubiquitin Ligase (E3) Involved in the Regulation of Carbohydrate Metabolism Mol. Biol. Cell, August 1, 2008; 19(8): 3323 - 3333. [Abstract] [Full Text] [PDF]

				J. C. Game, M. S. Williamson, and C. Baccari X-Ray Survival Characteristics and Genetic Analysis for Nine Saccharomyces Deletion Mutants That Show Altered Radiation Sensitivity Genetics, January 1, 2005; 169(1): 51 - 63. [Abstract] [Full Text] [PDF]

				G.-C. Hung, C. R. Brown, A. B. Wolfe, J. Liu, and H.-L. Chiang Degradation of the Gluconeogenic Enzymes Fructose-1,6-bisphosphatase and Malate Dehydrogenase Is Mediated by Distinct Proteolytic Pathways and Signaling Events J. Biol. Chem., November 19, 2004; 279(47): 49138 - 49150. [Abstract] [Full Text] [PDF]

				J. Onodera and Y. Ohsumi Ald6p Is a Preferred Target for Autophagy in Yeast, Saccharomyces cerevisiae J. Biol. Chem., April 16, 2004; 279(16): 16071 - 16076. [Abstract] [Full Text] [PDF]

				D.-Y. Cui, C. R. Brown, and H.-L. Chiang The Type 1 Phosphatase Reg1p-Glc7p Is Required for the Glucose-induced Degradation of Fructose-1,6-bisphosphatase in the Vacuole J. Biol. Chem., March 12, 2004; 279(11): 9713 - 9724. [Abstract] [Full Text] [PDF]

				C. R. Brown, J. Liu, G.-C. Hung, D. Carter, D. Cui, and H.-L. Chiang The Vid Vesicle to Vacuole Trafficking Event Requires Components of the SNARE Membrane Fusion Machinery J. Biol. Chem., July 3, 2003; 278(28): 25688 - 25699. [Abstract] [Full Text] [PDF]

				J. Regelmann, T. Schule, F. S. Josupeit, J. Horak, M. Rose, K.-D. Entian, M. Thumm, and D. H. Wolf Catabolite Degradation of Fructose-1,6-bisphosphatase in the Yeast Saccharomyces cerevisiae: A Genome-wide Screen Identifies Eight Novel GID Genes and Indicates the Existence of Two Degradation Pathways Mol. Biol. Cell, April 1, 2003; 14(4): 1652 - 1663. [Abstract] [Full Text] [PDF]

				C. R. Brown, J. A. McCann, G. G.-C. Hung, C. P. Elco, and H.-L. Chiang Vid22p, a novel plasma membrane protein, is required for the fructose-1,6-bisphosphatase degradation pathway J. Cell Sci., January 2, 2002; 115(3): 655 - 666. [Abstract] [Full Text] [PDF]

				C. R. Brown, D.-Y. Cui, G. G.-C. Hung, and H.-L. Chiang Cyclophilin A Mediates Vid22p Function in the Import of Fructose-1,6-bisphosphatase into Vid Vesicles J. Biol. Chem., December 14, 2001; 276(51): 48017 - 48026. [Abstract] [Full Text] [PDF]

				B. Gajewska, J. Kaminska, A. Jesionowska, N. C. Martin, A. K. Hopper, and T. Zoladek WW Domains of Rsp5p Define Different Functions: Determination of Roles in Fluid Phase and Uracil Permease Endocytosis in Saccharomyces cerevisiae Genetics, January 1, 2001; 157(1): 91 - 101. [Abstract] [Full Text]

				C. R. Brown, J. A. McCann, and H.-L. Chiang The Heat Shock Protein Ssa2p Is Required for Import of Fructose-1,6-Bisphosphatase into Vid Vesicles J. Cell Biol., July 11, 2000; 150(1): 65 - 76. [Abstract] [Full Text] [PDF]

				M. Horst, E. C. Knecht, and P. V. Schu Import into and Degradation of Cytosolic Proteins by Isolated Yeast Vacuoles Mol. Biol. Cell, September 1, 1999; 10(9): 2879 - 2889. [Abstract] [Full Text]

				J. M. Gancedo Yeast Carbon Catabolite Repression Microbiol. Mol. Biol. Rev., June 1, 1998; 62(2): 334 - 361. [Abstract] [Full Text] [PDF]

				D. J. Klionsky Nonclassical Protein Sorting to the Yeast Vacuole J. Biol. Chem., May 1, 1998; 273(18): 10807 - 10810. [Full Text] [PDF]

				M.-C. Chiang and H.-L. Chiang Vid24p, a Novel Protein Localized to the Fructose-1,6-bisphosphatase-containing Vesicles, Regulates Targeting of Fructose-1,6-bisphosphatase from the Vesicles to the Vacuole for Degradation J. Cell Biol., March 23, 1998; 140(6): 1347 - 1356. [Abstract] [Full Text] [PDF]

				N. J. Bryant and T. H. Stevens Vacuole Biogenesis in Saccharomyces cerevisiae: Protein Transport Pathways to the Yeast Vacuole Microbiol. Mol. Biol. Rev., March 1, 1998; 62(1): 230 - 247. [Abstract] [Full Text] [PDF]

				H.-L. Shieh and H.-L. Chiang In Vitro Reconstitution of Glucose-induced Targeting of Fructose-1,6-bisphosphatase into the Vacuole in Semi-intact Yeast Cells J. Biol. Chem., February 6, 1998; 273(6): 3381 - 3387. [Abstract] [Full Text] [PDF]

				P.-H. Huang and H.-L. Chiang Identification of Novel Vesicles in the Cytosol to Vacuole Protein Degradation Pathway J. Cell Biol., February 24, 1997; 136(4): 803 - 810. [Abstract] [Full Text] [PDF]

				H.-L. Shieh, Y. Chen, C. R. Brown, and H.-L. Chiang Biochemical Analysis of Fructose-1,6-bisphosphatase Import into Vacuole Import and Degradation Vesicles Reveals a Role for UBC1 in Vesicle Biogenesis J. Biol. Chem., March 23, 2001; 276(13): 10398 - 10406. [Abstract] [Full Text] [PDF]

				C. Andrei-Selmer, A. Knuppel, C. Satyanarayana, C. Heese, and P. V. Schu A New Class of Mutants Deficient in Dodecamerization of Aminopeptidase 1 and Vacuolar Transport J. Biol. Chem., April 6, 2001; 276(15): 11606 - 11614. [Abstract] [Full Text] [PDF]