Online citations, reference lists, and bibliographies.
← Back to Search

Aut5/Cvt17p, A Putative Lipase Essential For Disintegration Of Autophagic Bodies Inside The Vacuole.

U. D. Epple, I. Suriapranata, E. Eskelinen, M. Thumm
Published 2001 · Biology, Medicine

Cite This
Download PDF
Analyze on Scholarcy
Share
Selective disintegration of membrane-enclosed autophagic bodies is a feature of eukaryotic cells not studied in detail. Using a Saccharomyces cerevisiae mutant defective in autophagic-body breakdown, we identified and characterized Aut5p, a glycosylated integral membrane protein. Site-directed mutagenesis demonstrated the relevance of its putative lipase active-site motif for autophagic-body breakdown. aut5Delta cells show reduced protein turnover during starvation and are defective in maturation of proaminopeptidase I. Most recently, by means of the latter phenotype, Aut5p was independently identified as Cvt17p. In this study we additionally checked for effects on vacuolar acidification and detected mature vacuolar proteases, both of which are prerequisites for autophagic-body lysis. Furthermore, biologically active hemagglutinin-tagged Aut5p (Aut5-Ha) localizes to the endoplasmic reticulum (nuclear envelope) and is targeted to the vacuolar lumen independent of autophagy. In pep4Delta cells immunogold electron microscopy located Aut5-Ha at approximately 50-nm-diameter intravacuolar vesicles. Characteristic missorting in vps class E and fab1Delta cells, which affects the multivesicular body (MVB) pathway, suggests vacuolar targeting of Aut5-Ha similar to that of the MVB pathway. In agreement with localization of Aut5-Ha at intravacuolar vesicles in pep4Delta cells and the lack of vacuolar Aut5-Ha in wild-type cells, our pulse-chase experiments clearly indicated that Aut5-Ha degradation with 50 to 70 min of half-life is dependent on vacuolar proteinase A.
This paper references
10.1016/0378-1119(87)90232-0
A Saccharomyces cerevisiae genomic plasmid bank based on a centromere-containing shuttle vector.
M. Rose (1987)
10.1083/JCB.119.2.287
Aminopeptidase I of Saccharomyces cerevisiae is localized to the vacuole independent of the secretory pathway
D. Klionsky (1992)
10.1074/jbc.271.30.17621
Genetic and Phenotypic Overlap between Autophagy and the Cytoplasm to Vacuole Protein Targeting Pathway*
T. M. Harding (1996)
Current Protocols in Molecular Biology (New York: Greene Publishing Associates and Wiley-Interscience). Host-Range Shuttle System for Gene Insertion into the Chromosomes of Gram-negative Bacteria.
F. M. Ausubel (1988)
10.1093/nar/24.13.2519
A new efficient gene disruption cassette for repeated use in budding yeast
U. Güldener (1996)
10.1016/S0074-7696(00)98005-7
Alternative protein sorting pathways.
J. Kim (2000)
10.1111/j.1432-1033.1992.tb17118.x
Biogenesis of the yeast vacuole (lysosome). Proteinase yscB contributes molecularly and kinetically to vacuolar hydrolase-precursor maturation.
H. Hirsch (1992)
10.1083/JCB.124.6.903
Ultrastructural analysis of the autophagic process in yeast: detection of autophagosomes and their characterization
M. Baba (1994)
10.1002/YEA.320101310
New heterologous modules for classical or PCR‐based gene disruptions in Saccharomyces cerevisiae
A. Wach (1994)
The breakdown of autophagic vesicles inside the vacuole depends on Aut4p.
I. Suriapranata (2000)
10.1016/0962-8924(94)90069-8
Autophagy and related mechanisms of lysosome-mediated protein degradation.
W. Dunn (1994)
Lysosomal (vacuolar) proteinases of yeast are essential catalysts for protein degradation, differentiation, and cell survival.
U. Teichert (1989)
10.1002/1097-0029(20001215)51:6<563::AID-JEMT6>3.0.CO;2-8
Structure and function of the yeast vacuole and its role in autophagy
M. Thumm (2000)
10.1016/S0092-8674(00)81707-9
Fab1p PtdIns(3)P 5-Kinase Function Essential for Protein Sorting in the Multivesicular Body
G. Odorizzi (1998)
A Saccharomyces cerevisiae genomic plasmid bank based on a centromerecontaining shuttle vector. Gene 60:237–243
M. D. Rose (1987)
10.1083/JCB.119.2.301
Autophagy in yeast demonstrated with proteinase-deficient mutants and conditions for its induction
K. Takeshige (1992)
Biogenesis of the yeast vacuole (lysosome). The precursor forms of the soluble hydrolase carboxypeptidase yscS are associated with the vacuolar membrane.
D. Spormann (1992)
10.1038/35044114
A ubiquitin-like system mediates protein lipidation
Yoshinobu Ichimura (2000)
10.1002/(SICI)1097-0061(19980130)14:2<115::AID-YEA204>3.0.CO;2-2
Designer deletion strains derived from Saccharomyces cerevisiae S288C: A useful set of strains and plasmids for PCR‐mediated gene disruption and other applications
Carrie Baker Brachmann (1998)
10.1083/JCB.139.7.1687
Two Distinct Pathways for Targeting Proteins from the Cytoplasm to the Vacuole/Lysosome
M. Baba (1997)
10.1091/MBC.6.5.525
Novel PI(4)P 5-kinase homologue, Fab1p, essential for normal vacuole function and morphology in yeast.
A. Yamamoto (1995)
10.1016/0014-5793(94)00672-5
Isolation of autophagocytosis mutants of Saccharomyces cerevisiae
M. Thumm (1994)
10.1083/JCB.143.1.65
Fab1p Is Essential for PtdIns(3)P 5-Kinase Activity and the Maintenance of Vacuolar Size and Membrane Homeostasis
J. D. Gary (1998)
10.1128/JB.179.12.3875-3883.1997
AUT3, a serine/threonine kinase gene, is essential for autophagocytosis in Saccharomyces cerevisiae.
M. Straub (1997)
10.1007/BF00326437
The Saccharomyces cerevisiae HIS3 and LYS2 genes complement the Schizosaccharomyces pombe his5-303 and lys1-131 mutations, respectively: new selectable markers and new multi-purpose multicopy shuttle vectors, pSP3 and pSP4
G. Cottarel (2004)
10.1074/JBC.C000739200
Degradation of Lipid Vesicles in the Yeast Vacuole Requires Function of Cvt17, a Putative Lipase*
S. Teter (2001)
Structure and function of the yeast vacuole and its role in autophagy. Microsc
M. Thumm (2000)
10.1146/ANNUREV.CELLBIO.15.1.1
Vacuolar import of proteins and organelles from the cytoplasm.
D. Klionsky (1999)
10.1094/MPMI.1997.10.9.1106
Starvation-induced genes of the tomato pathogen Cladosporium fulvum are also induced during growth in planta.
M. Coleman (1997)
10.1083/JCB.150.6.1507
Tor-Mediated Induction of Autophagy via an Apg1 Protein Kinase Complex
Y. Kamada (2000)
10.1083/JCB.110.6.1923
Studies on the mechanisms of autophagy: formation of the autophagic vacuole
W. Dunn (1990)
10.1128/MCB.10.7.3737
Role of vacuolar acidification in protein sorting and zymogen activation: a genetic analysis of the yeast vacuolar proton-translocating ATPase.
C. Yamashiro (1990)
10.1016/S1569-2558(08)60457-9
From Proteasome to Lysosome: Studies on Yeast Demonstrate the Principles Of Protein Degradation in the Eukaryote Cell
M. Thumm (1998)
10.1073/PNAS.86.18.7027
Assay of vacuolar pH in yeast and identification of acidification-defective mutants.
R. Preston (1989)
10.1093/OXFORDJOURNALS.JBCHEM.A021592
Acidification of vacuoles is required for autophagic degradation in the yeast, Saccharomyces cerevisiae.
N. Nakamura (1997)
10.1016/S0968-0004(00)01543-7
Phosphoinositide signaling and the regulation of membrane trafficking in yeast.
G. Odorizzi (2000)
10.1083/JCB.152.3.519
Two Distinct Vps34 Phosphatidylinositol 3–Kinase Complexes Function in Autophagy and Carboxypeptidase Y Sorting inSaccharomyces cerevisiae
A. Kihara (2001)
VMA12 is essential for assembly of the vacuolar H(+)-ATPase subunits onto the vacuolar membrane in Saccharomyces cerevisiae.
R. Hirata (1993)
10.1128/JB.179.4.1068-1076.1997
AUT1, a gene essential for autophagocytosis in the yeast Saccharomyces cerevisiae.
M. Schlumpberger (1997)
10.1016/S1369-5274(99)00050-8
Bacterial replication in the host cell cytosol.
W. Goebel (2000)
10.1016/0076-6879(91)94047-G
Methods for studying the yeast vacuole.
C. J. Roberts (1991)
10.1128/MCB.15.11.5879
Endocytosis and vacuolar degradation of the plasma membrane-localized Pdr5 ATP-binding cassette multidrug transporter in Saccharomyces cerevisiae.
R. Egner (1995)
10.1093/emboj/17.17.4930
Phosphoinositide signaling and turnover: PtdIns(3)P, a regulator of membrane traffic, is transported to the vacuole and degraded by a process that requires lumenal vacuolar hydrolase activities
A. Wurmser (1998)
10.1093/emboj/17.13.3597
Aut2p and Aut7p, two novel microtubule‐associated proteins are essential for delivery of autophagic vesicles to the vacuole
T. Lang (1998)
10.1146/ANNUREV.BB.15.060186.001541
Identifying nonpolar transbilayer helices in amino acid sequences of membrane proteins.
D. Engelman (1986)



This paper is referenced by
10.3390/cells1030449
Role of Macroautophagy in Nutrient Homeostasis During Fungal Development and Pathogenesis
Yizhen Deng (2012)
10.1016/J.TCB.2004.07.014
Peroxisome turnover by micropexophagy: an autophagy-related process.
J. Farré (2004)
10.1007/s41745-016-0015-z
Multifaceted Housekeeping Functions of Autophagy
Sarika Chinchwadkar (2017)
10.1016/j.bbamcr.2008.08.003
The yeast lysosome-like vacuole: endpoint and crossroads.
S. C. Li (2009)
10.5772/17177
Biosensors for Monitoring Autophagy
Rod Devenish (2011)
CHARACTERIZATION OF YBR074 (PFF1), A CONSERVED VACUOLAR MEMBRANE METALLOPROTEASE FAMILY MEMBER
Karen A. Hecht (2013)
10.3389/fcell.2020.00221
Mitochondrial Fusion Machinery Specifically Involved in Energy Deprivation-Induced Autophagy
Choufei Wu (2020)
10.1242/jcs.01620
The molecular machinery of autophagy: unanswered questions
D. Klionsky (2005)
10.2323/JGAM.2017.08.001
Saccharomyces cerevisiae lipid droplet associated enzyme Ypr147cp shows both TAG lipase and ester hydrolase activities.
M. N. Kumar (2018)
Atg26 is involved in selective autophagy of the major coat protein Gag of the S. cerevisiae virus L-A
Peter Rube (2016)
Regulation of Mammalian Autophagy by Unc-51-Like Kinase 1
Hildegard I. D. Mack (2011)
10.1007/978-1-4614-5847-0_4
The Role of Autophagy in Drug Resistance and Potential for Therapeutic Targeting
R. Rangwala (2013)
10.1016/BS.IRCMB.2019.08.003
Molecular mechanisms of selective autophagy in Drosophila.
Raksha Gohel (2020)
10.1080/15548627.2015.1056969
A defect of the vacuolar putative lipase Atg15 accelerates degradation of lipid droplets through lipolysis
Y. Maéda (2015)
Autophagy and Vacuole Homeostasis
D. Mijaljica (2007)
10.1089/ars.2010.3762
Mitochondria autophagy in yeast.
T. Kanki (2011)
10.1091/MBC.E06-08-0663
Direct binding to Rsp5 mediates ubiquitin-independent sorting of Sna3 via the multivesicular body pathway.
M. McNatt (2007)
10.1186/s13578-020-00426-y
Synergies in exosomes and autophagy pathways for cellular homeostasis and metastasis of tumor cells
L. Salimi (2020)
10.1091/mbc.E17-08-0516
A newly characterized vacuolar serine carboxypeptidase, Atg42/Ybr139w, is required for normal vacuole function and the terminal steps of autophagy in the yeast Saccharomyces cerevisiae
Katherine R Parzych (2018)
10.1002/9781118845363.CH5
Protein Aggregation in Unicellular Eukaryotes
Marina Caldara (2014)
Effects of ghrelin on doxorubicin-induced toxicity in skeletal muscle
Pak Hung Yu (2013)
10.1007/s13238-010-0121-z
The late stage of autophagy: cellular events and molecular regulation
J. Tong (2010)
10.1016/j.bbamcr.2008.09.020
Principles of lysosomal membrane degradation: Cellular topology and biochemistry of lysosomal lipid degradation.
H. Schulze (2009)
10.1016/S0074-7696(03)32001-7
Dynamics of endosomal sorting.
N. Bishop (2003)
10.1089/ars.2010.3528
The role of macroautophagy in development of filamentous fungi.
M. Bartoszewska (2011)
10.3390/cells9051184
Closing the Gap: Membrane Contact Sites in the Regulation of Autophagy
Verena Kohler (2020)
10.1111/j.1365-3040.2010.02117.x
Devil inside: does plant programmed cell death involve the endomembrane system?
Jean-Luc Cacas (2010)
LSM1 AND RNY1: CLUES IN THE SEARCH FOR HOW RNA METABOLIC PATHWAYS CONTROL CANCER
Natalie Luhtala (2011)
10.15698/cst2019.05.186
Recent progress in the role of autophagy in neurological diseases
Tian Meng (2019)
10.1016/j.bbamcr.2015.01.005
Mitophagy in yeast: Molecular mechanisms and physiological role.
T. Kanki (2015)
10.7554/eLife.07736
The coordinated action of the MVB pathway and autophagy ensures cell survival during starvation
M. Müller (2015)
10.13130/morisi-federica_phd2016-01-11
AUTOPHAGY AND SKELETAL MUSCLE WASTING: EFFECTS ON SATELLITE CELLS POPULATION
F. Morisi (2016)
See more
Semantic Scholar Logo Some data provided by SemanticScholar