Human ATG4 autophagy proteases counteract attachment of ubiquitin-like LC3/GABARAP proteins to other cellular proteins.
ATG4B
Atg8
GABARAPL2
LC3ylation
autophagy
cysteine protease
deconjugation
deubiquitylation (deubiquitination)
post-translational modification
ubiquitin-conjugating enzyme (E2 enzyme)
Journal
The Journal of biological chemistry
ISSN: 1083-351X
Titre abrégé: J Biol Chem
Pays: United States
ID NLM: 2985121R
Informations de publication
Date de publication:
23 08 2019
23 08 2019
Historique:
received:
27
06
2019
pubmed:
19
7
2019
medline:
27
3
2020
entrez:
19
7
2019
Statut:
ppublish
Résumé
Microtubule-associated protein 1 light chain 3 α (LC3)/GABA type A receptor-associated protein (GABARAP) comprises a family of ubiquitin-like proteins involved in (macro)autophagy, an important intracellular degradation pathway that delivers cytoplasmic material to lysosomes via double-membrane vesicles called autophagosomes. The only currently known cellular molecules covalently modified by LC3/GABARAP are membrane phospholipids such as phosphatidylethanolamine in the autophagosome membrane. Autophagy-related 4 cysteine peptidase (ATG4) proteases process inactive pro-LC3/GABARAP before lipidation, and the same proteases can also deconjugate LC3/GABARAP from lipids. To determine whether LC3/GABARAP has other molecular targets, here we generated a pre-processed LC3B mutant (Q116P) that is resistant to ATG4-mediated deconjugation. Upon expression in human cells and when assessed by immunoblotting under reducing and denaturing conditions, deconjugation-resistant LC3B accumulated in multiple forms and at much higher molecular weights than free LC3B. We observed a similar accumulation when pre-processed versions of all mammalian LC3/GABARAP isoforms were expressed in ATG4-deficient cell lines, suggesting that LC3/GABARAP can attach also to other larger molecules. We identified ATG3, the E2-like enzyme involved in LC3/GABARAP lipidation, as one target of conjugation with multiple copies of LC3/GABARAP. We show that LC3B-ATG3 conjugates are distinct from the LC3B-ATG3 thioester intermediate formed before lipidation, and we biochemically demonstrate that ATG4B can cleave LC3B-ATG3 conjugates. Finally, we determined ATG3 residue Lys-243 as an LC3B modification site. Overall, we provide the first cellular evidence that mammalian LC3/GABARAP post-translationally modifies proteins akin to ubiquitination ("LC3ylation"), with ATG4 proteases acting like deubiquitinating enzymes to counteract this modification ("deLC3ylation").
Identifiants
pubmed: 31315929
pii: S0021-9258(20)30296-9
doi: 10.1074/jbc.AC119.009977
pmc: PMC6709618
doi:
Substances chimiques
Apoptosis Regulatory Proteins
0
Autophagy-Related Proteins
0
GABARAP protein, human
0
MAP1LC3A protein, human
0
Microtubule-Associated Proteins
0
Ubiquitins
0
ATG4B protein, human
EC 3.4.22.-
Cysteine Endopeptidases
EC 3.4.22.-
Banques de données
PDB
['2Z0E']
Types de publication
Journal Article
Research Support, Non-U.S. Gov't
Langues
eng
Sous-ensembles de citation
IM
Pagination
12610-12621Subventions
Organisme : Wellcome Trust
ID : 105604/Z/14/Z
Pays : United Kingdom
Organisme : Medical Research Council
ID : MR/M02492X/1
Pays : United Kingdom
Organisme : Wellcome Trust
Pays : United Kingdom
Organisme : Medical Research Council
ID : MC_EX_G0800785
Pays : United Kingdom
Organisme : Medical Research Council
ID : MC_U12266B
Pays : United Kingdom
Informations de copyright
© 2019 Agrotis et al.
Références
J Biol Chem. 2000 Oct 20;275(42):32966-73
pubmed: 10837468
J Cell Biol. 2000 Oct 16;151(2):263-76
pubmed: 11038174
EMBO J. 2000 Nov 1;19(21):5720-8
pubmed: 11060023
J Biol Chem. 2001 Jan 19;276(3):1701-6
pubmed: 11096062
Nature. 2000 Nov 23;408(6811):488-92
pubmed: 11100732
J Cell Biol. 2001 Feb 19;152(4):657-68
pubmed: 11266458
J Biol Chem. 2002 Apr 19;277(16):13739-44
pubmed: 11825910
J Biol Chem. 2002 May 24;277(21):18619-25
pubmed: 11897782
J Biol Chem. 2003 Apr 18;278(16):14053-8
pubmed: 12473658
J Biol Chem. 2003 Aug 1;278(31):29278-87
pubmed: 12740394
J Biol Chem. 2003 Dec 19;278(51):51841-50
pubmed: 14530254
J Cell Sci. 2004 Jun 1;117(Pt 13):2805-12
pubmed: 15169837
J Biol Chem. 2004 Aug 27;279(35):36268-76
pubmed: 15187094
Genome Res. 2004 Oct;14(10B):2128-35
pubmed: 15489335
J Biol Chem. 2006 Feb 10;281(6):3017-24
pubmed: 16303767
FEBS J. 2006 Jun;273(11):2553-62
pubmed: 16704426
Autophagy. 2007 Jan-Feb;3(1):48-50
pubmed: 17102583
J Biol Chem. 2007 Mar 16;282(11):8036-43
pubmed: 17227760
EMBO J. 2007 Apr 4;26(7):1749-60
pubmed: 17347651
Autophagy. 2007 Sep-Oct;3(5):452-60
pubmed: 17534139
J Biol Chem. 2007 Aug 17;282(33):24131-45
pubmed: 17580304
Autophagy. 2007 Nov-Dec;3(6):542-5
pubmed: 17611390
Cell. 2007 Jul 13;130(1):165-78
pubmed: 17632063
J Biol Chem. 2007 Dec 28;282(52):37298-302
pubmed: 17986448
Nature. 2007 Dec 20;450(7173):1253-7
pubmed: 18097414
Mol Biol Cell. 2008 May;19(5):2092-100
pubmed: 18321988
Genome Biol. 2008 Apr 03;9(4):R64
pubmed: 18387192
Mol Biol Cell. 2008 Aug;19(8):3290-8
pubmed: 18508918
Mol Biol Cell. 2008 Nov;19(11):4762-75
pubmed: 18768753
J Biol Chem. 2009 Mar 20;284(12):8023-32
pubmed: 19150980
FEBS Lett. 2009 Apr 2;583(7):1078-83
pubmed: 19285500
EMBO J. 2009 May 6;28(9):1341-50
pubmed: 19322194
Nat Rev Mol Cell Biol. 2009 May;10(5):319-31
pubmed: 19352404
Nat Rev Mol Cell Biol. 2009 Aug;10(8):550-63
pubmed: 19626045
Nature. 2010 Jul 1;466(7302):68-76
pubmed: 20562859
J Clin Invest. 2010 Jul;120(7):2331-44
pubmed: 20577052
Cell. 2010 Aug 20;142(4):590-600
pubmed: 20723759
J Biol Chem. 2011 Mar 4;286(9):7327-38
pubmed: 21177865
J Biol Chem. 2011 Mar 25;286(12):10238-47
pubmed: 21247896
Dev Cell. 2011 Apr 19;20(4):444-54
pubmed: 21497758
Mol Cell. 2011 Oct 21;44(2):325-40
pubmed: 21906983
EMBO J. 2011 Nov 08;30(23):4701-11
pubmed: 22068051
Autophagy. 2012 Feb 1;8(2):177-86
pubmed: 22240591
Oncogene. 2012 Oct 4;31(40):4397-408
pubmed: 22249245
Autophagy. 2012 May 1;8(5):780-93
pubmed: 22622160
Autophagy. 2012 Jun;8(6):883-92
pubmed: 22652539
PLoS One. 2013 Sep 10;8(9):e74222
pubmed: 24040206
Nat Protoc. 2013 Nov;8(11):2281-2308
pubmed: 24157548
Cell Rep. 2013 Nov 14;5(3):826-38
pubmed: 24210823
Nat Methods. 2014 May;11(5):585-92
pubmed: 24658140
Nat Struct Mol Biol. 2014 Apr;21(4):336-45
pubmed: 24699082
J Biol Chem. 2015 Mar 27;290(13):8146-53
pubmed: 25645919
Nat Cell Biol. 2015 Mar;17(3):300-10
pubmed: 25686249
Bioinformatics. 2015 Sep 15;31(18):3078-80
pubmed: 25979474
J Biol Chem. 2015 Oct 30;290(44):26549-61
pubmed: 26378241
J Cell Sci. 2016 Mar 1;129(5):875-80
pubmed: 26906419
Cell Res. 2016 Apr;26(4):423-40
pubmed: 27002219
Oncotarget. 2016 Jul 5;7(27):41203-41216
pubmed: 27256984
Cancer Cell. 2016 Aug 8;30(2):214-228
pubmed: 27478040
J Cell Biol. 2016 Dec 19;215(6):857-874
pubmed: 27864321
Annu Rev Biochem. 2017 Jun 20;86:193-224
pubmed: 28460188
EMBO J. 2017 Jul 3;36(13):1811-1836
pubmed: 28596378
Nat Commun. 2017 Aug 18;8(1):294
pubmed: 28821708
Nat Commun. 2017 Aug 18;8(1):295
pubmed: 28821724
Nat Commun. 2018 Jan 8;9(1):95
pubmed: 29311554
Nucleic Acids Res. 2018 Jul 2;46(W1):W296-W303
pubmed: 29788355
Autophagy. 2019 Jun;15(6):976-997
pubmed: 30661429
EMBO J. 1998 Jul 1;17(13):3597-607
pubmed: 9649430
Nature. 1998 Sep 24;395(6700):395-8
pubmed: 9759731
J Biol Chem. 1998 Dec 18;273(51):33889-92
pubmed: 9852036