Home LiteratureArticle Details
PMID: 18245858 Published · ppublish English Journal Article Research Support, U.S. Gov't, Non-P.H.S.

The ATG12-conjugating enzyme ATG10 Is essential for autophagic vesicle formation in Arabidopsis thaliana.

Genetics ·Vol. 178 ·No. 3 ·2008-03-00 ·Pages 1339-53

Phillips AR, Suttangkakul A, Vierstra RD

Abstract

Autophagy is an important intracellular recycling system in eukaryotes that utilizes small vesicles to traffic cytosolic proteins and organelles to the vacuole for breakdown. Vesicle formation requires the conjugation of the two ubiquitin-fold polypeptides ATG8 and ATG12 to phosphatidylethanolamine and the ATG5 protein, respectively. Using Arabidopsis thaliana mutants affecting the ATG5 target or the ATG7 E1 required to initiate ligation of both ATG8 and ATG12, we previously showed that the ATG8/12 conjugation pathways together are important when plants encounter nutrient stress and during senescence. To characterize the ATG12 conjugation pathway specifically, we characterized a null mutant eliminating the E2-conjugating enzyme ATG10 that, similar to plants missing ATG5 or ATG7, cannot form the ATG12-ATG5 conjugate. atg10-1 plants are hypersensitive to nitrogen and carbon starvation and initiate senescence and programmed cell death (PCD) more quickly than wild type, as indicated by elevated levels of senescence- and PCD-related mRNAs and proteins during carbon starvation. As detected with a GFP-ATG8a reporter, atg10-1 and atg5-1 mutant plants fail to accumulate autophagic bodies inside the vacuole. These results indicate that ATG10 is essential for ATG12 conjugation and that the ATG12-ATG5 conjugate is necessary to form autophagic vesicles and for the timely progression of senescence and PCD in plants.

MeSH Terms
Amino Acid Sequence Apoptosis/drug effects Arabidopsis/cytology,drug effects,enzymology,growth & development Arabidopsis Proteins/chemistry,metabolism Autophagy/drug effects Autophagy-Related Protein 12 Autophagy-Related Protein 5 Autophagy-Related Proteins Carbon/pharmacology DNA Fragmentation/drug effects Genes, Essential Genetic Complementation Test Molecular Sequence Data Mutant Proteins/isolation & purification Mutation/genetics Nitrogen/pharmacology Phenotype Phosphoric Monoester Hydrolases/metabolism Plant Leaves/cytology,drug effects RNA, Plant/metabolism Seedlings/cytology,drug effects,metabolism Transgenes
Chemicals
ATG10 protein, Arabidopsis ATG12 protein, Arabidopsis Arabidopsis Proteins Autophagy-Related Protein 12 Autophagy-Related Protein 5 Autophagy-Related Proteins Mutant Proteins RNA, Plant Carbon Atg5 protein, Arabidopsis Phosphoric Monoester Hydrolases Nitrogen
Authors & Affiliations
3 authors, click to expand affiliations / ORCID
Phillips Allison R
Department of Genetics, University of Wisconsin, Madison, Wisconsin 53706, USA.
Suttangkakul Anongpat
Vierstra Richard D
References (67)
67 references, click to expand
  1. Leaf senescence and starvation-induced chlorosis are accelerated by the disruption of an Arabidopsis autophagy gene.
    Plant Physiol. 2002 Jul;129(3):1181-93 PMID: 12114572
  2. The PET1-CMS mitochondrial mutation in sunflower is associated with premature programmed cell death and cytochrome c release.
    Plant Cell. 2001 Aug;13(8):1803-18 PMID: 11487694
  3. Inhibitory effect of modified bafilomycins and concanamycins on P- and V-type adenosinetriphosphatases.
    Biochemistry. 1993 Apr 20;32(15):3902-6 PMID: 8385991
  4. Transcription analysis of arabidopsis membrane transporters and hormone pathways during developmental and induced leaf senescence.
    Plant Physiol. 2006 Jun;141(2):776-92 PMID: 16603661
  5. Molecular dissection of autophagy: two ubiquitin-like systems.
    Nat Rev Mol Cell Biol. 2001 Mar;2(3):211-6 PMID: 11265251
  6. A plant vacuolar protease, VPE, mediates virus-induced hypersensitive cell death.
    Science. 2004 Aug 6;305(5685):855-8 PMID: 15297671
  7. Identification of a novel chloroplast protein AtNYE1 regulating chlorophyll degradation during leaf senescence in Arabidopsis.
    Plant Physiol. 2007 Jul;144(3):1429-41 PMID: 17468209
  8. Isolation and characterization of autophagy-defective mutants of Saccharomyces cerevisiae.
    FEBS Lett. 1993 Oct 25;333(1-2):169-74 PMID: 8224160
  9. Formation process of autophagosome is traced with Apg8/Aut7p in yeast.
    J Cell Biol. 1999 Oct 18;147(2):435-46 PMID: 10525546
  10. The APG8/12-activating enzyme APG7 is required for proper nutrient recycling and senescence in Arabidopsis thaliana.
    J Biol Chem. 2002 Sep 6;277(36):33105-14 PMID: 12070171
  11. Floral dip: a simplified method for Agrobacterium-mediated transformation of Arabidopsis thaliana.
    Plant J. 1998 Dec;16(6):735-43 PMID: 10069079
  12. A custom microarray analysis of gene expression during programmed cell death in Arabidopsis thaliana.
    Plant J. 2002 May;30(4):431-46 PMID: 12028573
  13. Plant autophagy--more than a starvation response.
    Curr Opin Plant Biol. 2007 Dec;10(6):587-93 PMID: 17702643
  14. Dexamethasone-induced apoptosis involves cleavage of DNA to large fragments prior to internucleosomal fragmentation.
    J Biol Chem. 1993 Feb 15;268(5):3037-9 PMID: 8428979
  15. AtATG18a is required for the formation of autophagosomes during nutrient stress and senescence in Arabidopsis thaliana.
    Plant J. 2005 May;42(4):535-46 PMID: 15860012
  16. Hierarchy of Atg proteins in pre-autophagosomal structure organization.
    Genes Cells. 2007 Feb;12(2):209-18 PMID: 17295840
  17. Temporal responses of transcripts, enzyme activities and metabolites after adding sucrose to carbon-deprived Arabidopsis seedlings.
    Plant J. 2007 Feb;49(3):463-91 PMID: 17217462
  18. A ubiquitin-like system mediates protein lipidation.
    Nature. 2000 Nov 23;408(6811):488-92 PMID: 11100732
  19. A senescence-associated gene of Arabidopsis thaliana is distinctively regulated during natural and artificially induced leaf senescence.
    Plant Mol Biol. 1996 Feb;30(4):739-54 PMID: 8624406
  20. The ubiquitin 26S proteasome proteolytic pathway.
    Annu Rev Plant Biol. 2004;55:555-90 PMID: 15377232
  21. Development by self-digestion: molecular mechanisms and biological functions of autophagy.
    Dev Cell. 2004 Apr;6(4):463-77 PMID: 15068787
  22. Isolation and characterization of yeast mutants in the cytoplasm to vacuole protein targeting pathway.
    J Cell Biol. 1995 Nov;131(3):591-602 PMID: 7593182
  23. The gain-of-function Arabidopsis acd6 mutant reveals novel regulation and function of the salicylic acid signaling pathway in controlling cell death, defenses, and cell growth.
    Plant Cell. 1999 Sep;11(9):1695-708 PMID: 10488236
  24. Interrelationships among Atg proteins during autophagy in Saccharomyces cerevisiae.
    Yeast. 2004 Sep;21(12):1057-65 PMID: 15449304
  25. Arabidopsis AtBECLIN 1/AtAtg6/AtVps30 is essential for pollen germination and plant development.
    Cell Res. 2007 Mar;17(3):249-63 PMID: 17339883
  26. A comparison of the expression patterns of several senescence-associated genes in response to stress and hormone treatment.
    Plant Mol Biol. 1998 Jun;37(3):455-69 PMID: 9617813
  27. A unique mechanism for protein processing and degradation in Arabidopsis thaliana.
    Proc Natl Acad Sci U S A. 2003 Jun 10;100(12):7389-94 PMID: 12773619
  28. Genome-wide insertional mutagenesis of Arabidopsis thaliana.
    Science. 2003 Aug 1;301(5633):653-7 PMID: 12893945
  29. A protein conjugation system essential for autophagy.
    Nature. 1998 Sep 24;395(6700):395-8 PMID: 9759731
  30. Atg7-dependent autophagy promotes neuronal health, stress tolerance, and longevity but is dispensable for metamorphosis in Drosophila.
    Genes Dev. 2007 Dec 1;21(23):3061-6 PMID: 18056421
  31. Isolation of autophagocytosis mutants of Saccharomyces cerevisiae.
    FEBS Lett. 1994 Aug 1;349(2):275-80 PMID: 8050581
  32. Visualization of autophagy in Arabidopsis using the fluorescent dye monodansylcadaverine and a GFP-AtATG8e fusion protein.
    Plant J. 2005 May;42(4):598-608 PMID: 15860017
  33. Apg10p, a novel protein-conjugating enzyme essential for autophagy in yeast.
    EMBO J. 1999 Oct 1;18(19):5234-41 PMID: 10508157
  34. 2,4-Dichlorophenoxybutyric acid-resistant mutants of Arabidopsis have defects in glyoxysomal fatty acid beta-oxidation.
    Plant Cell. 1998 Feb;10(2):183-95 PMID: 9490742
  35. Constitutive expression of CIR1 (RVE2) affects several circadian-regulated processes and seed germination in Arabidopsis.
    Plant J. 2007 Aug;51(3):512-25 PMID: 17587236
  36. Programmed cell death: a way of life for plants.
    Proc Natl Acad Sci U S A. 1996 Oct 29;93(22):12094-7 PMID: 8901538
  37. Transcriptome profiling of the response of Arabidopsis suspension culture cells to Suc starvation.
    Plant Physiol. 2004 Aug;135(4):2330-47 PMID: 15310832
  38. Gateway-compatible vectors for plant functional genomics and proteomics.
    Plant J. 2006 Feb;45(4):616-29 PMID: 16441352
  39. Vacuolar processing enzyme is up-regulated in the lytic vacuoles of vegetative tissues during senescence and under various stressed conditions.
    Plant J. 1999 Jul;19(1):43-53 PMID: 10417725
  40. Autophagy and plant innate immunity.
    Cell Microbiol. 2006 Jun;8(6):899-906 PMID: 16681833
  41. A light-entrained circadian clock controls transcription of several plant genes.
    EMBO J. 1988 Dec 1;7(12):3635-42 PMID: 3208743
  42. Autophagy and cell-death proteases in plants: two wheels of a funeral cart.
    Autophagy. 2007 Mar-Apr;3(2):136-8 PMID: 17204839
  43. Apg16p is required for the function of the Apg12p-Apg5p conjugate in the yeast autophagy pathway.
    EMBO J. 1999 Jul 15;18(14):3888-96 PMID: 10406794
  44. Are diverse signalling pathways integrated in the regulation of arabidopsis antioxidant defence gene expression in response to excess excitation energy?
    Philos Trans R Soc Lond B Biol Sci. 2000 Oct 29;355(1402):1531-40 PMID: 11128006
  45. The crystal structure of plant ATG12 and its biological implication in autophagy.
    Autophagy. 2005 Jul;1(2):119-26 PMID: 16874047
  46. An Arabidopsis homolog of yeast ATG6/VPS30 is essential for pollen germination.
    Plant Physiol. 2007 Mar;143(3):1132-9 PMID: 17259285
  47. In vivo analysis of autophagy in response to nutrient starvation using transgenic mice expressing a fluorescent autophagosome marker.
    Mol Biol Cell. 2004 Mar;15(3):1101-11 PMID: 14699058
  48. Autophagic recycling: lessons from yeast help define the process in plants.
    Curr Opin Plant Biol. 2005 Apr;8(2):165-73 PMID: 15752997
  49. The mouse APG10 homologue, an E2-like enzyme for Apg12p conjugation, facilitates MAP-LC3 modification.
    J Biol Chem. 2003 Oct 10;278(41):39517-26 PMID: 12890687
  50. Cytokinin growth responses in Arabidopsis involve the 26S proteasome subunit RPN12.
    Plant Cell. 2002 Jan;14(1):17-32 PMID: 11826296
  51. Reactive oxygen species are essential for autophagy and specifically regulate the activity of Atg4.
    EMBO J. 2007 Apr 4;26(7):1749-60 PMID: 17347651
  52. Programmed Cell Death in Plants.
    Plant Cell. 1997 Jul;9(7):1157-1168 PMID: 12237381
  53. Formation of the approximately 350-kDa Apg12-Apg5.Apg16 multimeric complex, mediated by Apg16 oligomerization, is essential for autophagy in yeast.
    J Biol Chem. 2002 May 24;277(21):18619-25 PMID: 11897782
  54. Processing of ATG8s, ubiquitin-like proteins, and their deconjugation by ATG4s are essential for plant autophagy.
    Plant Cell. 2004 Nov;16(11):2967-83 PMID: 15494556
  55. The pre-autophagosomal structure organized by concerted functions of APG genes is essential for autophagosome formation.
    EMBO J. 2001 Nov 1;20(21):5971-81 PMID: 11689437
  56. Senescence is induced in individually darkened Arabidopsis leaves, but inhibited in whole darkened plants.
    Plant Physiol. 2001 Nov;127(3):876-86 PMID: 11706170
  57. Mouse Apg10 as an Apg12-conjugating enzyme: analysis by the conjugation-mediated yeast two-hybrid method.
    FEBS Lett. 2002 Dec 18;532(3):450-4 PMID: 12482611
  58. The autophagosomal-lysosomal compartment in programmed cell death.
    Cell Death Differ. 2001 Jun;8(6):569-81 PMID: 11536007
  59. Many ways to exit? Cell death categories in plants.
    Trends Plant Sci. 2005 Mar;10(3):117-22 PMID: 15749469
  60. The VTI family of SNARE proteins is necessary for plant viability and mediates different protein transport pathways.
    Plant Cell. 2003 Dec;15(12):2885-99 PMID: 14615598
  61. Degradation of oxidized proteins by autophagy during oxidative stress in Arabidopsis.
    Plant Physiol. 2007 Jan;143(1):291-9 PMID: 17098847
  62. Autophagy regulates programmed cell death during the plant innate immune response.
    Cell. 2005 May 20;121(4):567-577 PMID: 15907470
  63. The reversible modification regulates the membrane-binding state of Apg8/Aut7 essential for autophagy and the cytoplasm to vacuole targeting pathway.
    J Cell Biol. 2000 Oct 16;151(2):263-76 PMID: 11038174
  64. Autophagic nutrient recycling in Arabidopsis directed by the ATG8 and ATG12 conjugation pathways.
    Plant Physiol. 2005 Aug;138(4):2097-110 PMID: 16040659
  65. Starvation-induced expression of autophagy-related genes in Arabidopsis.
    Biol Cell. 2006 Jan;98(1):53-67 PMID: 16354162
  66. The autophagy-associated Atg8 gene family operates both under favourable growth conditions and under starvation stresses in Arabidopsis plants.
    J Exp Bot. 2005 Nov;56(421):2839-49 PMID: 16157655
  67. Autophagy: from phenomenology to molecular understanding in less than a decade.
    Nat Rev Mol Cell Biol. 2007 Nov;8(11):931-7 PMID: 17712358
Article Info
Journal
Genetics
Abbr.
Genetics
ISSN
0016-6731
DOI
10.1534/genetics.107.086199
Published
2008-03-00
Epub
2008-00-03
Pages
1339-53
Language
English
Region
United States
NLM ID
0374636
PMCID
PMC2278079
Subset
IM
External Links
PubMed source DOI: 10.1534/genetics.107.086199 PMC Full Text View journal details
Analysis Services
Analysis Services

Contact

No. 2 Wenbo Road, Zhangqiu District, Jinan, Shandong

Qilu Normal University · Genelibs Bioinformatics Lab

750 Shunhua Rd, Jinan

2F, Bldg F, University Science Park

Tel: 0531-88819269

WeChat Official Account

Follow our WeChat subscription account for real-time updates and the latest in medical and biological research.


Business Email

E-mail: [email protected]

Copyright © 2026.Genelibs Jinan Honghui Biotechnology Co., Ltd. 鲁ICP备20007159号-2