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Cited by 64Autophagy in Disease and TherapyReproductive tract infections researchBacterial Genetics and Biotechnology

Autophagy-independent function of MAP-LC3 during intracellular propagation of Chlamydia trachomatis

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Al-Younes, Hesham M., Al-Zeer, Munir A., Khalil, Hany, Gussmann, Joscha, Karlas, Alexander, Machuy, Nikolaus, Brinkmann, Volker, Braun, Peter R. +1 more·2011·Autophagy
DOI 10.4161/auto.7.8.15597
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Abstract

Microtubule-associated protein 1 (MAP1) light chain 3 (LC3) has proven useful as autophagosomal marker in studies on the interaction between pathogens and the host autophagic machinery. However, the function of LC3 is known to extend above and beyond its role in autophagosome formation. We previously reported that intrinsic LC3 is associated with the intracellular Chlamydia trachomatis inclusion in human epithelial cells. Here we show that LC3, most likely the cytoplasmic nonlipidated form, interacts with the C. trachomatis inclusion as a microtubule-associated protein rather than an autophagosome-associated component. In contrast, N-terminally GFP-tagged LC3 exclusively targets autophagosomes rather than chlamydial inclusions. Immunofluorescence analysis revealed an association of LC3 and MAP1 subunits A and B with the inclusion as early as 18 h post infection. Inclusion-bound LC3 was connected with the microtubular network. Depolymerization of the microtubular architecture disrupted the association of LC3/MAP1s with the inclusion. Furthermore, siRNA-mediated silencing of the MAP1 and LC3 proteins revealed their essential function in the intracellular growth of C. trachomatis. Interestingly, defective autophagy remarkably enhanced chlamydial growth, suggesting a suppressive effect of the autophagic machinery on bacterial development. However, depletion of LC3 in autophagy-deficient cells noticeably reduced chlamydial propagation. Thus, our findings demonstrate a new function for LC3, distinct from autophagy, in intracellular bacterial pathogenesis.

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  • LC3 can associate with other cellular components including microtubules and the osteoclast ruffled border observed in bone resorption.

    “LC3 can also associate with other cellular components including microtubules [19] and the osteoclast ruffled border observed in bone resorption [20].”

    From LC3-Associated Phagocytosis (LAP): Connections with Host Autophagy (2012) · cites this paper as [19]

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Research works printed but not linked: 58

  1. Hammerschlag MR. Chlamydia trachomatis and Chlamydia pneumoniae infections in children and ado- lescents. Pediatri Rev 2004; 25:43-51.
  2. Fields KA, Hackstadt T. The chlamydial inclusion: escape from the endocytic pathway. Annu Rev Cell Dev Bio 2002; 18:221-45.
  3. van Ooij C, Apodaca G, Engel J. Characterization of the Chlamydia trachomatis vacuole and its interaction with the host endocytic pathway in HeLa cells. Infect Immun 1997; 65:758-66.
  4. Al-Younes HM, Rudel T, Brinkmann V, Szczepek AJ, Meyer TF. Low iron availability modulates the course of Chlamydia pneumoniae infection. Cell Microbiol 2001; 3:427-37.
  5. Al-Younes HM, Rudel T, Meyer TF. Characterization and intracellular trafficking pattern of vacuoles contain- ing Chlamydia pneumoniae in human epithelial cells. Cell Microbiol 1999; 1:237-47.
  6. Rejman Lipinski A, Heymann J, Meissner C, Karlas A, Brinkmann V, Meyer TF, et al. Rab6 and Rab11 regulate Chlamydia trachomatis development and gol- gin-84-dependent Golgi fragmentation. PLoS Pathog 2009; 5:1000615.
  7. Heuer D, Lipinski AR, Machuy N, Karlas A, Wehrens A, Siedler F, et al. Chlamydia causes fragmentation of the Golgi compartment to ensure reproduction. Nature 2009; 457:731-5.
  8. Heinzen R, Scidmore M, Rockey D, Hackstadt T. Differential interaction with endocytic and exocyt- ic pathways distinguish parasitophorous vacuoles of Coxiella burnetii and Chlamydia trachomatis. Infect Immun 1996; 64:796-809.
  9. Mizushima N, Yamamoto A, Hatano M, Kobayashi Y, Kabeya Y, Suzuki K, et al. Dissection of autophagosome formation using Apg5-deficient mouse embryonic stem cells. J Cell Biol 2001; 152:657-68.
  10. Kirkegaard K, Taylor MP, Jackson WT. Cellular auto- phagy: surrender, avoidance and subversion by micro- organisms. Nat Rev Micro 2004; 2:301-14.
  11. Kabeya Y, Mizushima N, Ueno T, Yamamoto A, Kirisako T, Noda T, et al. LC3, a mammalian homo- logue of yeast Apg8p, is localized in autophagosome membranes after processing. EMBO J 2000; 19:5720-8.
  12. Al-Younes HM, Brinkmann V, Meyer TF. Interaction of Chlamydia trachomatis serovar L2 with the host auto- phagic pathway. Infect Immun 2004; 72:4751-62.
  13. Amano A, Nakagawa I, Yoshimori T. Autophagy in innate immunity against intracellular bacteria. J Biochem 2006; 140:161-6.
  14. Gutierrez MG, Master SS, Singh SB, Taylor GA, Colombo MI, Deretic V. Autophagy is a defense mechanism inhibiting BCG and Mycobacterium tuber- culosis survival in infected macrophages. Cell 2004; 119:753-66.
  15. Nakagawa I, Amano A, Mizushima N, Yamamoto A, Yamaguchi H, Kamimoto T, et al. Autophagy defends cells against invading group A Streptococcus. Science 2004; 306:1037-40.
  16. Romano PS, Gutierrez MG, Berón W, Rabinovitch M, Colombo MI. The autophagic pathway is actively modulated by phase II Coxiella burnetii to efficiently replicate in the host cell. Cell Microbiol 2007; 9:891-909.
  17. Mizushima N, Yamamoto A, Matsui M, Yoshimori T, Ohsumi Y. In vivo analysis of autophagy in response to nutrient starvation using transgenic mice expressing a fluorescent autophagosome marker. Mol Biol Cell 2004; 15:1101-11.
  18. Mann S, Hammarback J. Molecular characterization of light chain 3. A microtubule binding subunit of MAP1A and MAP1B. J Biol Chem 1994; 269:11492-7.
  19. Kuznetsov SA, Gelfand VI. 18 kDa microtubule- associated protein: identification as a new light chain (LC-3) of microtubule-associated protein 1 (MAP-1). FEBS Lett 1987; 212:145-8.
  20. Mann S, Hammarback J. Gene localization and devel- opmental expression of light chain 3: A common sub- unit of microtubule-associated protein 1A (MAP1A) and MAP1B. J Neurosci Res 1996; 43:535-44.
  21. Seidenbecher CI, Landwehr M, Smalla KH, Kreutz M, Dieterich DC, Zuschratter W, et al. Caldendrin but not calmodulin binds to light chain 3 of MAP1A/B: An association with the microtubule cytoskeleton highlighting exclusive binding partners for neuronal Ca 2+ -sensor proteins. J Mol Biol 2004; 336:957-70.
  22. Zhou B, Rabinovitch M. Microtubule involvement in translational regulation of fibronectin expression by light chain 3 of microtubule-associated protein 1 in vascular smooth muscle cells. Circ Res 1998; 83:481-9.
  23. Takemura R, Okabe S, Umeyama T, Kanai Y, Cowan N, Hirokawa N. Increased microtubule stability and alpha tubulin acetylation in cells transfected with microtubule-associated proteins MAP1B, MAP2 or tau. J Cell Sci 1992; 103:953-64.
  24. Halpain S, Dehmelt L. The MAP1 family of microtu- bule-associated proteins. Genome Biol 2006; 7:224.
  25. Faller EM, Villeneuve TS, Brown DL. MAP1a associ- ated light chain 3 increases microtubule stability by suppressing microtubule dynamics. Mol Cell Neurosci 2009; 41:85-93.
  26. Kabeya Y, Mizushima N, Yamamoto A, Oshitani- Okamoto S, Ohsumi Y, Yoshimori T. LC3, GABARAP and GATE16 localize to autophagosomal membrane depending on form-II formation. J Cell Sci 2004; 117:2805-12.
  27. Tanida I, Ueno T, Kominami E. 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 2004; 279:47704-10.
  28. He H, Dang Y, Dai F, Guo Z, Wu J, She X, et al. Post- translational modifications of three members of the human MAP1LC3 family and detection of a novel type of modification for MAP1LC3B. J Biol Chem 2003; 278:29278-87.
  29. Hackstadt T, Rockey D, Heinzen R, Scidmore M. Chlamydia trachomatis interrupts an exocytic pathway to acquire endogenously synthesized sphingomyelin in transit from the Golgi apparatus to the plasma mem- brane. EMBO J 1996; 15:964-77.
  30. Clausen JD, Christiansen G, Holst HU, Birkelund S. Chlamydia trachomatis utilizes the host cell micro- tubule network during early events of infection. Mol Microbiol 1997; 25:441-9.
  31. Hemelaar J, Lelyveld VS, Kessler BM, Ploegh HL. A single protease, Apg4B, is specific for the autopha- gy-related ubiquitin-like proteins GATE-16, MAP1- LC3, GABARAP and Apg8L. J Biol Chem 2003; 278:51841-50.
  32. Nakatogawa H, Suzuki K, Kamada Y, Ohsumi Y. Dynamics and diversity in autophagy mechanisms: lessons from yeast. Nat Rev Mol Cell Biol 2009; 10:458-67.
  33. Behrends C, Sowa ME, Gygi SP, Harper JW. Network organization of the human autophagy system. Nature 2010; 466:68-84.
  34. Münz C. Enhancing immunity through autophagy. Annu Rev Immunol 2009; 27:423-49.
  35. Escalante-Ochoa C, Ducatelle R, Haesebrouck F. Optimal development of Chlamydophila psittaci in L929 fibroblast and BGM epithelial cells requires the participation of microfilaments and microtubule-motor proteins. Microb Pathogenesis 2000; 28:321-33.
  36. Kumar Y, Valdivia RH. Actin and intermediate fila- ments stabilize the Chlamydia trachomatis vacuole by forming dynamic structural scaffolds. Cell Host Microbe 2008; 4:159-69.
  37. Gurumurthy RK, Maurer AP, Machuy N, Hess S, Pleissner KP, Schuchhardt J, et al. A loss-of-function screen reveals Ras-and Raf-independent MEK-ERK signaling during Chlamydia trachomatis infection. Sci Signal 2010; 3:11.
  38. Machuy N, Thiede B, Rajalingam K, Dimmler C, Thieck O, Meyer TF, et al. A global approach combin- ing proteome analysis and phenotypic screening with RNA interference yields novel apoptosis regulators. Mol Cell Proteomics 2005; 4:44-55.
  39. Wiznerowicz M, Trono D. Conditional suppression of cellular genes: Lentivirus vector-mediated drug- inducible RNA interference. J Virol 2003; 77:8957-61.
  40. Kouno T, Mizuguchi M, Tanida I, Ueno T, Kanematsu T, Mori Y, et al. Solution structure of microtubule- associated protein light chain 3 and identification of its functional subdomains. J Biol Chem 2005; 280:24610-7.
  41. Wu J, Dang Y, Su W, Liu C, Ma H, Shan Y, et al. Molecular cloning and characterization of rat LC3A and LC3B-Two novel markers of autophagosome. Biochem Biophys Res Commun 2006; 339:437-42.
  42. Cann GM, Guignabert C, Ying L, Deshpande N, Bekker JM, Wang L, et al. Developmental expression of LC3α and β: Absence of fibronectin or autophagy phenotype in LC3β knockout mice. Devl Dynam 2008; 237:187-95.
  43. Chakrama FZ, Seguin-Py S, Le Grand JN, Fraichard A, Delage-Mourroux R, Despouy G, et al. GABARAPL1 (GEC1) associates with autophagic vesicles. Autophagy 2010; 6:495-505.
  44. Cabrera S, Marino G, Fernandez AF, Lopez-Otin C. Autophagy, proteases and the sense of balance. Autophagy 2010; 6:961-3.
  45. Schwarten M, Mohrluder J, Ma PX, Stoldt M, Thielmann Y, Stangler T, et al. Nix directly binds to GABARAP A possible crosstalk between apoptosis and autophagy. Autophagy 2009; 5:690-8.
  46. Klebig C, Seitz S, Arnold W, Deutschmann N, Pacyna-Gengelbach M, Scherneck S, et al. Characterization of gamma-aminobutyric acid type A receptor associated protein, a novel tumor suppressor, showing reduced expression in breast cancer. Cancer Res 2005; 65:394-400.
  47. Comes F, Matrone A, Lastella P, Nico B, Susca FC, Bagnulo R, et al. A novel cell type-specific role of p38 alpha in the control of autophagy and cell death in colorectal cancer cells. Cell Death Differ 2007; 14:693-702.
  48. Wang H, Olsen RW. Binding of the GABAA receptor- associated protein (GABARAP) to microtubules and microfilaments suggests involvement of the cytoskel- eton in GABARAPGABAA receptor interaction. J Neurochem 2000; 75:644-55.
  49. Mansuy V, Boireau W, Fraichard A, Schlick JL, Jouvenot M, Delage-Mourroux R. GEC1, a protein related to GABARAP, interacts with tubulin and GABAA receptor. Biochem Biophys Res Commun 2004; 325:639-48.
  50. Scidmore MA, Hackstadt T. Mammalian 14-3-3β associates with the Chlamydia trachomatis inclusion membrane via its interaction with IncG. Mol Microbiol 2001; 39:1638-50.
  51. Mizushima N, Yoshimori T, Levine B. Methods in mam- malian autophagy research. Cell 2010; 140:313-26.
  52. Kuma A, Hatano M, Matsui M, Yamamoto A, Nakaya H, Yoshimori T, et al. The role of autophagy during the early neonatal starvation period. Nature 2004; 432:1032-6.
  53. Munafo DB, Colombo MI. A novel assay to study auto- phagy: regulation of autophagosome vacuole size by amino acid deprivation. J Cell Sci 2001; 114:3619-29.
  54. Bampton E, Goemans C, Niranjan D, Mizushima N, Tolkovsky A. The dynamics of autophagy visualized in live cells: from autophagosome formation to fusion with endo/lysosomes. Autophagy 2005; 1:23-36.
  55. Klionsky D, Abeliovich H, Agostinis P, Agrawal D, Aliev G, Askew D, et al. Guidelines for the use of interpretation of assays for monitoring autophagy in higher eukaryotes. Autophagy 2008; 4:151-75.
  56. Niemann A, Takatsuki A, Elsässer HP. The lysoso- motropic agent monodansylcadaverine also acts as a solvent polarity probe. J Histochem Cytochem 2000; 48:251-8.
  57. Lajoie P, Guay G, Dennis JW, Nabi IR. The lipid com- position of autophagic vacuoles regulates expression of multilamellar bodies. J Cell Sci 2005; 118:1991-2003.
  58. Beron W, Gutierrez MG, Rabinovitch M, Colombo MI. Coxiella burnetii localizes in a Rab7-labeled compartment with autophagic characteristics. Infect Immun 2002; 70:5816-21.

Cited by

Sorted by citations

Authors
Al-Younes, H. M., Al-Zeer, M. A., Khalil, H., Gussmann, J., Karlas, A., Machuy, N., Brinkmann, V., Braun, P. R., & Meyer, T. F.
Affiliations
  • University of Jordan (JO)
  • Max Planck Society (DE)
Corresponding author
Meyer, Thomas F.
ORCID
Venue
Autophagy
Publisher
Informa UK Limited
Published
2011-08
Volume
7
Issue
8
Pages
814-828
ISSN
  • 1554-8627 (ISSN-L)
  • 1554-8635
DOI
10.4161/auto.7.8.15597
OpenAlex
W1990490178
Cited by
64
References
0
Citations by year
2025: 3 · 2024: 4 · 2023: 7 · 2022: 6 · 2021: 5 · 2020: 3 · 2019: 6 · 2018: 2 · 2017: 4 · 2016: 5 · 2015: 5 · 2014: 4 · 2013: 3 · 2012: 7
FWCI
3.36
Type
journal article
Language
en
Open access
Locations
  • Autophagy (publishedVersion)
  • PubMed (publishedVersion)
  • Max Planck Institute for Plasma Physics (submittedVersion)
Funders
Deutsche Forschungsgemeinschaft
Topics
  • Health Sciences › Medicine › Epidemiology › Autophagy in Disease and Therapy
  • Life Sciences › Immunology and Microbiology › Microbiology › Reproductive tract infections research
  • Life Sciences › Biochemistry, Genetics and Molecular Biology › Genetics › Bacterial Genetics and Biotechnology
Keywords
LC3, Chlamydia trachomatis, autophagy-independent function, chlamydial inclusions, microtubule-associated proteins, MAP1A, MAP1B, microtubule cytoskeleton, intracellular bacterial growth, host-pathogen interaction
MeSH terms
Autophagy-Related Protein 5, Animals, Autophagy, Inclusion Bodies, Chlamydia trachomatis, HeLa Cells, Humans, Intracellular Membranes, Lipids, Microtubule-Associated Proteins, Microtubules, Phagosomes, Recombinant Fusion Proteins, Protein Biosynthesis, Protein Subunits, Protein Transport, RNA, Small Interfering, Intracellular Space, Green Fluorescent Proteins, Mice

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