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Autophagy In Tobacco BY-2 Cells Cultured Under Sucrose Starvation Conditions: Isolation Of The Autolysosome And Its Characterization.

Chihiro Takatsuka, Y. Inoue, Tomoya Higuchi, S. Hillmer, D. Robinson, Y. Moriyasu
Published 2011 · Biology, Medicine

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Tobacco culture cells carry out a large-scale degradation of intracellular proteins in order to survive under sucrose starvation conditions. We have previously suggested that this bulk degradation of cellular proteins is performed by autophagy, where autolysosomes formed de novo act as the major lytic compartments. The digestion process in autolysosomes can be retarded by addition of the cysteine protease inhibitor E-64c to the culture medium, resulting in the accumulation of autolysosomes. In the present study, we have investigated several properties of autolysosomes in tobacco cells. Electron microscopy showed that the autolysosomes contain osmiophilic particles, some of which resemble partially degraded mitochondria. It also revealed the presence of two kinds of autolysosome precursor structures; one resembled the isolation membrane and the other the autophagosome of mammalian cells. Immunofluorescence microscopy showed that autolysosomes contain acid phosphatase, in accordance with cytochemical enzyme analyses by light and electron microscopy in a previous study. Autolysosomes isolated by cell fractionation on Percoll gradients showed the localization of acid phosphatase, vacuolar H(+)-ATPase and cysteine protease. These results show that starvation-induced autophagy in tobacco cells follows a macroautophagic-type response similar to that described for other eukaryotes. However, our results indicate that, although the plant vacuole is often described as being equivalent to the lysosome of the animal cell, a new low pH lytic compartment-the autolysosome-also contributes to proteolytic degradation when tobacco cells are subjected to sucrose deprivation.
This paper references
10.4161/auto.2.2.2366
Protein Aggregates are Transported to Vacuoles by Macroautophagic Mechanism in Nutrient-Starved Plant Cells
K. Toyooka (2006)
10.1104/pp.011024
Leaf Senescence and Starvation-Induced Chlorosis Are Accelerated by the Disruption of an Arabidopsis Autophagy Gene1
Hideki Hanaoka (2002)
10.1093/PCP/PCJ013
Autophagy is not a main contributor to the degradation of phospholipids in tobacco cells cultured under sucrose starvation conditions.
Y. Inoue (2006)
10.1104/pp.111.4.1233
Autophagy in Tobacco Suspension-Cultured Cells in Response to Sucrose Starvation
Y. Moriyasu (1996)
10.1074/JBC.C000739200
Degradation of Lipid Vesicles in the Yeast Vacuole Requires Function of Cvt17, a Putative Lipase*
S. Teter (2001)
10.1038/227680A0
Cleavage of Structural Proteins during the Assembly of the Head of Bacteriophage T4
U. Laemmli (1970)
10.1105/tpc.104.025395
Processing of ATG8s, Ubiquitin-Like Proteins, and Their Deconjugation by ATG4s Are Essential for Plant Autophagy
K. Yoshimoto (2004)
10.1111/J.1365-313X.2004.02282.X
Essential role of the V-ATPase in male gametophyte development.
Jan Dettmer (2005)
10.1016/0962-8924(94)90069-8
Autophagy and related mechanisms of lysosome-mediated protein degradation.
W. Dunn (1994)
10.1111/j.1749-6632.1964.tb14213.x
DISC ELECTROPHORESIS – II METHOD AND APPLICATION TO HUMAN SERUM PROTEINS *
B. J. Davis (1964)
10.1104/pp.105.060673
Autophagic Nutrient Recycling in Arabidopsis Directed by the ATG8 and ATG12 Conjugation Pathways1
Allison R Thompson (2005)
10.1093/PCP/PCH105
Contribution of the plasma membrane and central vacuole in the formation of autolysosomes in cultured tobacco cells.
Kanako Yano (2004)
10.1093/PCP/PCG100
Alpha tonoplast intrinsic protein is specifically associated with vacuole membrane involved in an autophagic process.
Y. Moriyasu (2003)
10.1083/JCB.119.2.301
Autophagy in yeast demonstrated with proteinase-deficient mutants and conditions for its induction
K. Takeshige (1992)
10.4161/auto.2092
Autophagy in Development and Stress Responses of Plants
D. Bassham (2006)
10.1042/BC20040516
Starvation‐induced expression of autophagy‐related genes in Arabidopsis
T. Rose (2006)
10.1016/S1534-5807(03)00296-X
A unified nomenclature for yeast autophagy-related genes.
D. Klionsky (2003)
10.1016/0014-4800(85)90020-6
Uptake--microautophagy--and degradation of exogenous proteins by isolated rat liver lysosomes. Effects of pH, ATP, and inhibitors of proteolysis.
J. Ahlberg (1985)
10.1083/JCB.136.1.61
The Autophagic and Endocytic Pathways Converge at the Nascent Autophagic Vacuoles
W. Liou (1997)
10.1146/ANNUREV.CELLBIO.15.1.1
Vacuolar import of proteins and organelles from the cytoplasm.
D. Klionsky (1999)
10.1126/SCIENCE.290.5497.1717
Autophagy as a regulated pathway of cellular degradation.
D. Klionsky (2000)
10.1073/PNAS.75.2.852
Cytochemical studies on GERL, provacuoles, and vacuoles in root meristematic cells of Euphorbia.
F. Marty (1978)
10.1016/S0076-6879(08)03232-1
Use of protease inhibitors for detecting autophagy in plants.
Y. Moriyasu (2008)
10.1016/J.PBI.2007.06.006
Plant autophagy--more than a starvation response.
D. Bassham (2007)
10.1104/pp.106.092106
Degradation of Oxidized Proteins by Autophagy during Oxidative Stress in Arabidopsis1[W][OA]
Y. Xiong (2006)
10.1016/0076-6879(91)94043-C
Immunofluorescence methods for yeast.
J. Pringle (1991)
10.1093/PCP/PCL031
AtATG genes, homologs of yeast autophagy genes, are involved in constitutive autophagy in Arabidopsis root tip cells.
Y. Inoue (2006)
10.1007/BF01279880
Vacuole biogenesis and protein transport to the plant vacuole: A comparison with the yeast vacuole and the mammalian lysosome
D. Robinson (2005)
10.1007/s00709-004-0070-6
The V-ATPase inhibitors concanamycin A and bafilomycin A lead to Golgi swelling in tobacco BY-2 cells
D. Robinson (2004)
10.1105/tpc.105.037978
Vacuolar H+-ATPase Activity Is Required for Endocytic and Secretory Trafficking in Arabidopsis[W]
Jan Dettmer (2006)
10.1016/0003-9861(82)90504-5
6-substituted purines: a novel class of inhibitors of endogenous protein degradation in isolated rat hepatocytes.
P. B. Gordon (1982)
10.1111/J.1365-313X.2005.02349.X
Endosomal proteases facilitate the fusion of endosomes with vacuoles at the final step of the endocytotic pathway.
Kenji Yamada (2005)
10.1105/tpc.109.072637
Endocytic and Secretory Traffic in Arabidopsis Merge in the Trans-Golgi Network/Early Endosome, an Independent and Highly Dynamic Organelle[W]
Corrado Viotti (2010)
10.1104/pp.108.122770
Mobilization of Rubisco and Stroma-Localized Fluorescent Proteins of Chloroplasts to the Vacuole by an ATG Gene-Dependent Autophagic Process1[W][OA]
H. Ishida (2008)
10.4161/auto.1.1.1270
Maturation of Autophagic Vacuoles in Mammalian Cells
E. Eskelinen (2005)
10.1083/JCB.147.2.435
Formation Process of Autophagosome Is Traced with Apg8/Aut7p in Yeast
T. Kirisako (1999)
10.1016/j.semcdb.2010.03.009
Regulation of macroautophagy in Saccharomyces cerevisiae.
Yuko Inoue (2010)
Sequestration of cytoplasmic enzymes in an autophagic vacuole-lysosomal system induced by injection of leupeptin.
E. Kominami (1983)
10.4161/auto.3678
Methods for Monitoring Autophagy from Yeast to Human
D. Klionsky (2007)
10.1083/JCB.141.3.625
Peroxisome Degradation by Microautophagy in Pichia pastoris: Identification of Specific Steps and Morphological Intermediates
Y. Sakai (1998)
10.4161/auto.6845
Does bafilomycin A1 block the fusion of autophagosomes with lysosomes?
D. Klionsky (2008)
10.1093/PCP/PCH031
3-methyladenine inhibits autophagy in tobacco culture cells under sucrose starvation conditions.
Chihiro Takatsuka (2004)
10.1083/JCB.133.6.1251
Ultrastructural and biochemical characterization of autophagy in higher plant cells subjected to carbon deprivation: control by the supply of mitochondria with respiratory substrates
S. Aubert (1996)
10.1105/tpc.11.4.601
Protein Storage Bodies and Vacuoles
E. Herman (1999)
Divergent modes of autophagy in the methylotrophic yeast Pichia pastoris.
D. Tuttle (1995)
10.1007/BF03036132
Autophagy in plants
S. Kwon (2009)
10.1111/J.1365-313X.2005.02396.X
Visualization of autophagy in Arabidopsis using the fluorescent dye monodansylcadaverine and a GFP-AtATG8e fusion protein.
Anthony L Contento (2005)
10.1016/0003-2697(84)90553-0
Fluorescein isothiocyanate-labeled casein assay for proteolytic enzymes.
S. Twining (1984)
10.1073/PNAS.79.6.1889
3-Methyladenine: specific inhibitor of autophagic/lysosomal protein degradation in isolated rat hepatocytes.
P. Seglen (1982)



This paper is referenced by
10.1016/j.aca.2012.09.041
Review on recent advances in the analysis of isolated organelles.
C. Satori (2012)
10.3390/cells9040887
Microautophagy in Plants: Consideration of Its Molecular Mechanism
K. Sieńko (2020)
10.1016/j.ymeth.2014.09.003
Methods for analysis of autophagy in plants.
D. Bassham (2015)
10.1371/journal.pone.0045673
Different Degree in Proteasome Malfunction Has Various Effects on Root Growth Possibly through Preventing Cell Division and Promoting Autophagic Vacuolization
X. Sheng (2012)
10.1007/s10059-012-0098-y
Genes for plant Autophagy: Functions and interactions
S. Kim (2012)
10.1093/pcp/pcs099
Beginning to understand autophagy, an intracellular self-degradation system in plants.
K. Yoshimoto (2012)
10.1105/tpc.113.118307
A BAR-Domain Protein SH3P2, Which Binds to Phosphatidylinositol 3-Phosphate and ATG8, Regulates Autophagosome Formation in Arabidopsis[C][W]
Xiaohong Zhuang (2013)
10.4161/auto.26275
Ultrastructure of autophagy in plant cells
W. G. van Doorn (2013)
10.1021/acs.analchem.6b02920
Plant Cell Wall-Penetrable, Redox-Responsive Silica Nanoprobe for the Imaging of Starvation-Induced Vesicle Trafficking.
Xin-Chun Huang (2016)
10.1007/978-1-4939-6533-5_12
Isolation of Autolysosomes from Tobacco BY-2 Cells.
Chihiro Takatsuka (2017)
10.3390/ijms21010194
Autophagy-Like Cell Death Regulates Hydrogen Peroxide and Calcium Ion Distribution in Xa3/Xa26-Mediated Resistance to Xanthomonas oryzae pv. oryzae
Jianbo Cao (2019)
10.1039/c8fo02295j
Biological properties of plant-derived extracellular vesicles.
S. Rome (2019)
10.1080/15592324.2015.1082699
Dissection of autophagy in tobacco BY-2 cells under sucrose starvation conditions using the vacuolar H+-ATPase inhibitor concanamycin A and the autophagy-related protein Atg8
Kanako Yano (2015)
10.1242/jcs.093559
Structure and function of endosomes in plant cells
Anthony L Contento (2012)
10.1104/pp.114.254078
Establishment of Monitoring Methods for Autophagy in Rice Reveals Autophagic Recycling of Chloroplasts and Root Plastids during Energy Limitation1[OPEN]
M. Izumi (2015)
10.1111/j.1744-7909.2012.01178.x
What to eat: evidence for selective autophagy in plants.
Brice E. Floyd (2012)
10.1038/s41598-019-45470-y
Nitric oxide and ROS mediate autophagy and regulate Alternaria alternata toxin-induced cell death in tobacco BY-2 cells
Abhishek Sadhu (2019)
10.4161/psb.6.12.18297
Detecting autophagy in Arabidopsis roots by membrane-permeable cysteine protease inhibitor E-64d and endocytosis tracer FM4–64
Yuumi Oh-ye (2011)
10.1007/978-3-319-21033-9_11
To Live or Die: Autophagy in Plants
Brice E. Floyd (2015)
10.1093/pcp/pct129
Cell wall polysaccharides are mislocalized to the Vacuole in echidna mutants.
H. McFarlane (2013)
10.1080/09168451.2020.1756736
Sucrose starvation induces the degradation of proteins in trans-Golgi network and secretory vesicle cluster in tobacco BY-2 cells
Yamato Oda (2020)
10.1007/s11627-014-9622-4
Impact of carbon and phosphate starvation on growth and programmed cell death of maritime pine suspension cells
H. Azevedo (2014)
10.1016/J.ENVEXPBOT.2019.05.005
Cell death signaling and morphology in chemical-treated tobacco BY-2 suspension cultured cells
E. Iakimova (2019)
10.1093/pcp/pcu041
Assessment and optimization of autophagy monitoring methods in Arabidopsis roots indicate direct fusion of autophagosomes with vacuoles.
E. A. Merkulova (2014)
10.3390/cells7010005
Multiscale and Multimodal Approaches to Study Autophagy in Model Plants
J. Marion (2018)
10.1111/ppl.12124
Cadmium-induced cell death in BY-2 cell culture starts with vacuolization of cytoplasm and terminates with necrosis.
J. Kutík (2014)
10.1007/978-4-431-54941-3_5
Vacuoles and Storage Organelles
T. Noguchi (2014)
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