Regulation of ATG4B Stability by RNF5 Limits Basal Levels of Autophagy and Influences Susceptibility to Bacterial Infection

English version České info

Autophagy is the mechanism by which cytoplasmic components and organelles are degraded by the lysosomal machinery in response to diverse stimuli including nutrient deprivation, intracellular pathogens, and multiple forms of cellular stress. Here, we show that the membrane-associated E3 ligase RNF5 regulates basal levels of autophagy by controlling the stability of a select pool of the cysteine protease ATG4B. RNF5 controls the membranal fraction of ATG4B and limits LC3 (ATG8) processing, which is required for phagophore and autophagosome formation. The association of ATG4B with—and regulation of its ubiquitination and stability by—RNF5 is seen primarily under normal growth conditions. Processing of LC3 forms, appearance of LC3-positive puncta, and p62 expression are higher in RNF5^−/− MEF. RNF5 mutant, which retains its E3 ligase activity but does not associate with ATG4B, no longer affects LC3 puncta. Further, increased puncta seen in RNF5^−/− using WT but not LC3 mutant, which bypasses ATG4B processing, substantiates the role of RNF5 in early phases of LC3 processing and autophagy. Similarly, RNF-5 inactivation in Caenorhabditis elegans increases the level of LGG-1/LC3::GFP puncta. RNF5^−/− mice are more resistant to group A Streptococcus infection, associated with increased autophagosomes and more efficient bacterial clearance by RNF5^−/− macrophages. Collectively, the RNF5-mediated control of membranalATG4B reveals a novel layer in the regulation of LC3 processing and autophagy.

Vyšlo v časopise: Regulation of ATG4B Stability by RNF5 Limits Basal Levels of Autophagy and Influences Susceptibility to Bacterial Infection. PLoS Genet 8(10): e32767. doi:10.1371/journal.pgen.1003007
Kategorie: Research Article
prolekare.web.journal.doi_sk: https://doi.org/10.1371/journal.pgen.1003007

Souhrn

Zdroje

1. KunduM, ThompsonCB (2008) Autophagy: basic principles and relevance to disease. Annu Rev Pathol 3 : 427–455.

2. HeC, KlionskyDJ (2009) Regulation mechanisms and signaling pathways of autophagy. Annu Rev Genet 43 : 67–93.

3. MizushimaN, LevineB (2010) Autophagy in mammalian development and differentiation. Nat Cell Biol 12 : 823–830.

4. LevineB (2007) Cell biology: autophagy and cancer. Nature 446 : 745–747.

5. LevineB, MizushimaN, VirginHW (2011) Autophagy in immunity and inflammation. Nature 469 : 323–335.

6. LevineB, KroemerG (2008) Autophagy in the pathogenesis of disease. Cell 132 : 27–42.

7. MizushimaN, LevineB, CuervoAM, KlionskyDJ (2008) Autophagy fights disease through cellular self-digestion. Nature 451 : 1069–1075.

8. DereticV, LevineB (2009) Autophagy, immunity, and microbial adaptations. Cell Host Microbe 5 : 527–549.

9. NakagawaI, AmanoA, MizushimaN, YamamotoA, YamaguchiH, et al. (2004) Autophagy defends cells against invading group A Streptococcus. Science 306 : 1037–1040.

10. MizushimaN, YoshimoriT, LevineB (2010) Methods in mammalian autophagy research. Cell 140 : 313–326.

11. MizushimaN (2007) Autophagy: process and function. Genes Dev 21 : 2861–2873.

12. KlionskyDJ, CreggJM, DunnWAJr, EmrSD, SakaiY, et al. (2003) A unified nomenclature for yeast autophagy-related genes. Dev Cell 5 : 539–545.

13. GengJ, KlionskyDJ (2008) The Atg8 and Atg12 ubiquitin-like conjugation systems in macroautophagy. ‘Protein modifications: beyond the usual suspects’ review series. EMBO Rep 9 : 859–864.

14. OhsumiY (2001) Molecular dissection of autophagy: two ubiquitin-like systems. Nat Rev Mol Cell Biol 2 : 211–216.

15. TanidaI, UenoT, KominamiE (2004) Human light chain 3/MAP1LC3B is cleaved at its carboxyl-terminal Met121 to expose Gly120 for lipidation and targeting to autophagosomal membranes. J Biol Chem 279 : 47704–47710.

16. KabeyaY, MizushimaN, UenoT, YamamotoA, KirisakoT, et al. (2000) LC3, a mammalian homologue of yeast Apg8p, is localized in autophagosome membranes after processing. Embo J 19 : 5720–5728.

17. KabeyaY, MizushimaN, YamamotoA, Oshitani-OkamotoS, OhsumiY, et al. (2004) LC3, GABARAP and GATE16 localize to autophagosomal membrane depending on form-II formation. J Cell Sci 117 : 2805–2812.

18. IchimuraY, KirisakoT, TakaoT, SatomiY, ShimonishiY, et al. (2000) A ubiquitin-like system mediates protein lipidation. Nature 408 : 488–492.

19. KirisakoT, IchimuraY, OkadaH, KabeyaY, MizushimaN, et al. (2000) The reversible modification regulates the membrane-binding state of Apg8/Aut7 essential for autophagy and the cytoplasm to vacuole targeting pathway. J Cell Biol 151 : 263–276.

20. TanidaI, SouYS, EzakiJ, Minematsu-IkeguchiN, UenoT, et al. (2004) HsAtg4B/HsApg4B/autophagin-1 cleaves the carboxyl termini of three human Atg8 homologues and delipidates microtubule-associated protein light chain 3 -⁠ and GABAA receptor-associated protein-phospholipid conjugates. J Biol Chem 279 : 36268–36276.

21. NakatogawaH, IshiiJ, AsaiE, OhsumiY (2012) Atg4 recycles inappropriately lipidated Atg8 to promote autophagosome biogenesis. Autophagy 8.

22. MarinoG, UriaJA, PuenteXS, QuesadaV, BordalloJ, et al. (2003) Human autophagins, a family of cysteine proteinases potentially implicated in cell degradation by autophagy. J Biol Chem 278 : 3671–3678.

23. HemelaarJ, LelyveldVS, KesslerBM, PloeghHL (2003) A single protease, Apg4B, is specific for the autophagy-related ubiquitin-like proteins GATE-16, MAP1-LC3, GABARAP, and Apg8L. J Biol Chem 278 : 51841–51850.

24. LiM, HouY, WangJ, ChenX, ShaoZM, et al. (2011) Kinetics comparisons of mammalian Atg4 homologues indicate selective preferences toward diverse Atg8 substrates. J Biol Chem 286 : 7327–7338.

25. BetinVM, LaneJD (2009) Caspase cleavage of Atg4D stimulates GABARAP-L1 processing and triggers mitochondrial targeting and apoptosis. J Cell Sci 122 : 2554–2566.

26. MarinoG, Salvador-MontoliuN, FueyoA, KnechtE, MizushimaN, et al. (2007) Tissue-specific autophagy alterations and increased tumorigenesis in mice deficient in Atg4C/autophagin-3. J Biol Chem 282 : 18573–18583.

27. MarinoG, FernandezAF, CabreraS, LundbergYW, CabanillasR, et al. (2010) Autophagy is essential for mouse sense of balance. J Clin Invest 120 : 2331–2344.

28. LuZ, LuoRZ, LuY, ZhangX, YuQ, et al. (2008) The tumor suppressor gene ARHI regulates autophagy and tumor dormancy in human ovarian cancer cells. J Clin Invest 118 : 3917–3929.

29. ChenZH, KimHP, SciurbaFC, LeeSJ, Feghali-BostwickC, et al. (2008) Egr-1 regulates autophagy in cigarette smoke-induced chronic obstructive pulmonary disease. PLoS ONE 3: e3316 doi:10.1371/journal.pone.0003316.

30. FujitaN, NodaT, YoshimoriT (2009) Atg4B(C74A) hampers autophagosome closure: a useful protein for inhibiting autophagy. Autophagy 5 : 88–89.

31. FujitaN, Hayashi-NishinoM, FukumotoH, OmoriH, YamamotoA, et al. (2008) An Atg4B mutant hampers the lipidation of LC3 paralogues and causes defects in autophagosome closure. Mol Biol Cell 19 : 4651–4659.

32. BrodayL, KolotuevI, DidierC, BhoumikA, PodbilewiczB, et al. (2004) The LIM domain protein UNC-95 is required for the assembly of muscle attachment structures and is regulated by the RING finger protein RNF-5 in C. elegans. J Cell Biol 165 : 857–867.

33. Zaidel-BarR, MillerS, KaminskyR, BrodayL (2010) Molting-specific downregulation of C. elegans body-wall muscle attachment sites: the role of RNF-5 E3 ligase. Biochem Biophys Res Commun 395 : 509–514.

34. DidierC, BrodayL, BhoumikA, IsraeliS, TakahashiS, et al. (2003) RNF5, a RING finger protein that regulates cell motility by targeting paxillin ubiquitination and altered localization. Mol Cell Biol 23 : 5331–5345.

35. DelaunayA, BrombergKD, HayashiY, MirabellaM, BurchD, et al. (2008) The ER-bound RING finger protein 5 (RNF5/RMA1) causes degenerative myopathy in transgenic mice and is deregulated in inclusion body myositis. PLoS ONE 3: e1609 doi:10.1371/journal.pone.0001609.

36. TcherpakovM, DelaunayA, TothJ, KadoyaT, PetroskiMD, et al. (2009) Regulation of endoplasmic reticulum-associated degradation by RNF5-dependent ubiquitination of JNK-associated membrane protein (JAMP). J Biol Chem 284 : 12099–12109.

37. YoungerJM, ChenL, RenHY, RosserMF, TurnbullEL, et al. (2006) Sequential quality-control checkpoints triage misfolded cystic fibrosis transmembrane conductance regulator. Cell 126 : 571–582.

38. BrombergKD, KlugerHM, DelaunayA, AbbasS, DiVitoKA, et al. (2007) Increased expression of the E3 ubiquitin ligase RNF5 is associated with decreased survival in breast cancer. Cancer Res 67 : 8172–8179.

39. ZhongB, ZhangL, LeiC, LiY, MaoAP, et al. (2009) The ubiquitin ligase RNF5 regulates antiviral responses by mediating degradation of the adaptor protein MITA. Immunity 30 : 397–407.

40. ZhongB, ZhangY, TanB, LiuTT, WangYY, et al. (2010) The E3 ubiquitin ligase RNF5 targets virus-induced signaling adaptor for ubiquitination and degradation. J Immunol 184 : 6249–6255.

41. ZhangY, HigashideW, DaiS, ShermanDM, ZhouD (2005) Recognition and ubiquitination of Salmonella type III effector SopA by a ubiquitin E3 ligase, HsRMA1. J Biol Chem 280 : 38682–38688.

42. Scherz-ShouvalR, ShvetsE, FassE, ShorerH, GilL, et al. (2007) Reactive oxygen species are essential for autophagy and specifically regulate the activity of Atg4. Embo J 26 : 1749–1760.

43. ShuCW, DragM, BekesM, ZhaiD, SalvesenGS, et al. (2010) Synthetic substrates for measuring activity of autophagy proteases: autophagins (Atg4). Autophagy 6 : 936–947.

44. KettelerR, SeedB (2008) Quantitation of autophagy by luciferase release assay. Autophagy 4 : 801–806.

45. KettelerR, SunZ, KovacsKF, HeWW, SeedB (2008) A pathway sensor for genome-wide screens of intracellular proteolytic cleavage. Genome Biol 9: R64.

46. KimuraS, NodaT, YoshimoriT (2007) Dissection of the autophagosome maturation process by a novel reporter protein, tandem fluorescent-tagged LC3. Autophagy 3 : 452–460.

47. CalfonM, ZengH, UranoF, TillJH, HubbardSR, et al. (2002) IRE1 couples endoplasmic reticulum load to secretory capacity by processing the XBP-1 mRNA. Nature 415 : 92–96.

48. YoungAR, ChanEY, HuXW, KochlR, CrawshawSG, et al. (2006) Starvation and ULK1-dependent cycling of mammalian Atg9 between the TGN and endosomes. J Cell Sci 119 : 3888–3900.

49. Yla-AnttilaP, VihinenH, JokitaloE, EskelinenEL (2009) 3D tomography reveals connections between the phagophore and endoplasmic reticulum. Autophagy 5 : 1180–1185.

50. ToozeSA, YoshimoriT (2010) The origin of the autophagosomal membrane. Nat Cell Biol 12 : 831–835.

51. StarrT, ChildR, WehrlyTD, HansenB, HwangS, et al. (2012) Selective subversion of autophagy complexes facilitates completion of the Brucella intracellular cycle. Cell Host Microbe 11 : 33–45.

52. HermanAM, MoussaCE (2011) The ubiquitin ligase parkin modulates the execution of autophagy. Autophagy 7 : 919–921.

53. LamarkT, KirkinV, DikicI, JohansenT (2009) NBR1 and p62 as cargo receptors for selective autophagy of ubiquitinated targets. Cell Cycle 8 : 1986–1990.

54. GanleyIG, WongPM, GammohN, JiangX (2011) Distinct autophagosomal-lysosomal fusion mechanism revealed by thapsigargin-induced autophagy arrest. Mol Cell 42 : 731–743.

55. JiaK, ThomasC, AkbarM, SunQ, Adams-HuetB, et al. (2009) Autophagy genes protect against Salmonella typhimurium infection and mediate insulin signaling-regulated pathogen resistance. Proc Natl Acad Sci U S A 106 : 14564–14569.

56. BirminghamCL, SmithAC, BakowskiMA, YoshimoriT, BrumellJH (2006) Autophagy controls Salmonella infection in response to damage to the Salmonella-containing vacuole. J Biol Chem 281 : 11374–11383.

57. DereticV, MasterS, SinghS (2008) Autophagy gives a nod and a wink to the inflammasome and Paneth cells in Crohn's disease. Dev Cell 15 : 641–642.

58. TimmerAM, TimmerJC, PenceMA, HsuLC, GhochaniM, et al. (2009) Streptolysin O promotes group A Streptococcus immune evasion by accelerated macrophage apoptosis. J Biol Chem 284 : 862–871.

59. De VriesL, ElenkoE, McCafferyJM, FischerT, HublerL, et al. (1998) RGS-GAIP, a GTPase-activating protein for Galphai heterotrimeric G proteins, is located on clathrin-coated vesicles. Mol Biol Cell 9 : 1123–1134.

60. AladzsityI, TothML, SigmondT, SzaboE, BicsakB, et al. (2007) Autophagy genes unc-51 and bec-1 are required for normal cell size in Caenorhabditis elegans. Genetics 177 : 655–660.

61. MelendezA, TalloczyZ, SeamanM, EskelinenEL, HallDH, et al. (2003) Autophagy genes are essential for dauer development and life-span extension in C. elegans. Science 301 : 1387–1391.

62. DaromA, Bening-Abu-ShachU, BrodayL (2010) RNF-121 is an endoplasmic reticulum-membrane E3 ubiquitin ligase involved in the regulation of beta-integrin. Mol Biol Cell 21 : 1788–1798.

63. FraserAG, KamathRS, ZipperlenP, Martinez-CamposM, SohrmannM, et al. (2000) Functional genomic analysis of C. elegans chromosome I by systematic RNA interference. Nature 408 : 325–330.