Identification of a candidate therapeutic autophagy-inducing peptide

Subjects

Abstract

The lysosomal degradation pathway of autophagy has a crucial role in defence against infection, neurodegenerative disorders, cancer and ageing. Accordingly, agents that induce autophagy may have broad therapeutic applications. One approach to developing such agents is to exploit autophagy manipulation strategies used by microbial virulence factors. Here we show that a peptide, Tat–beclin 1—derived from a region of the autophagy protein, beclin 1, which binds human immunodeficiency virus (HIV)-1 Nef—is a potent inducer of autophagy, and interacts with a newly identified negative regulator of autophagy, GAPR-1 (also called GLIPR2). Tat–beclin 1 decreases the accumulation of polyglutamine expansion protein aggregates and the replication of several pathogens (including HIV-1) in vitro, and reduces mortality in mice infected with chikungunya or West Nile virus. Thus, through the characterization of a domain of beclin 1 that interacts with HIV-1 Nef, we have developed an autophagy-inducing peptide that has potential efficacy in the treatment of human diseases.

Access options

Rent or Buy article

Get time limited or full article access on ReadCube.

from$8.99

All prices are NET prices.

Figure 1: Tat–beclin 1 peptide induces autophagy in vitro.
Figure 2: Tat–beclin 1 peptide binds to GAPR-1, a beclin 1-interacting protein.
Figure 3: Tat–beclin 1 peptide decreases aggregates of a polyglutamine expansion protein and has anti-infective activity.
Figure 4: Tat–beclin 1 peptide induces autophagy and exerts antiviral activity in vivo.

References

  1. 1

    Levine, B. & Kroemer, G. Autophagy in the pathogenesis of disease. Cell 132, 27–42 (2008)

    CAS  PubMed  PubMed Central  Google Scholar 

  2. 2

    Mizushima, N., Levine, B., Cuervo, A. M. & Klionsky, D. J. Autophagy fights disease through cellular self-digestion. Nature 451, 1069–1075 (2008)

    ADS  CAS  PubMed  PubMed Central  Google Scholar 

  3. 3

    Rubinsztein, D. C., Codogno, P. & Levine, B. Autophagy modulation as a potential therapeutic target for diverse diseases. Nature Rev. Drug Discov. 11, 709–730 (2012)

    CAS  Google Scholar 

  4. 4

    Kihara, A., Kabeya, Y., Ohsumi, Y. & Yoshimori, T. Beclin-phosphatidylinositol 3-kinase complex functions at the trans-Golgi network. EMBO Rep. 2, 330–335 (2001)

    CAS  PubMed  PubMed Central  Google Scholar 

  5. 5

    Kyei, G. B. et al. Autophagy pathway intersects with HIV-1 biosynthesis and regulates viral yields in macrophages. J. Cell Biol. 186, 255–268 (2009)

    CAS  PubMed  PubMed Central  Google Scholar 

  6. 6

    Furuya, N., Yu, J., Byfield, M., Pattingre, S. & Levine, B. The evolutionarily conserved domain of Beclin 1 is required for Vps34 binding, autophagy and tumor suppressor function. Autophagy 1, 46–52 (2005)

    CAS  PubMed  Google Scholar 

  7. 7

    Huang, W. et al. Crystal structure and biochemical analyses reveal Beclin 1 as a novel membrane binding protein. Cell Res. 22, 473–489 (2012)

    CAS  PubMed  PubMed Central  Google Scholar 

  8. 8

    Liang, X. H. et al. Induction of autophagy and inhibition of tumorigenesis by beclin 1. Nature 402, 672–676 (1999)

    ADS  CAS  PubMed  PubMed Central  Google Scholar 

  9. 9

    Mizushima, N., Yoshimori, T. & Levine, B. Methods in mammalian autophagy research. Cell 140, 313–326 (2010)

    CAS  PubMed  PubMed Central  Google Scholar 

  10. 10

    van den Berg, A. & Dowdy, S. F. Protein transduction domain delivery of therapeutic macromolecules. Curr. Opin. Biotechnol. 22, 888–893 (2011)

    CAS  PubMed  Google Scholar 

  11. 11

    Bayer, P. et al. Structural studies of HIV-1 Tat protein. J. Mol. Biol. 247, 529–535 (1995)

    CAS  PubMed  Google Scholar 

  12. 12

    Lee, J. S. et al. FLIP-mediated autophagy regulation in cell death control. Nature Cell Biol. 11, 1355–1362 (2009)

    CAS  PubMed  Google Scholar 

  13. 13

    Orvedahl, A. O. et al. Autophagy protects against Sindbis virus infection of the central nervous system. Cell Host Microbe 7, 115–127 (2010)

    CAS  PubMed  PubMed Central  Google Scholar 

  14. 14

    Eberle, H. B. et al. Identification and characterization of a novel human plant pathogenesis-related protein that localizes to lipid-enriched microdomains in the Golgi complex. J. Cell Sci. 115, 827–838 (2002)

    CAS  PubMed  Google Scholar 

  15. 15

    Polson, H. E. et al. Mammalian Atg18 (WIPI2) localizes to omegasome-anchored phagophores and positively regulates LC3 lipidation. Autophagy 6, 506–522 (2010)

    CAS  PubMed  Google Scholar 

  16. 16

    Harris, H. & Rubinsztein, D. C. Control of autophagy as a therapy for neurodegenerative disease. Nature Rev. Neurol. 8, 108–117 (2012)

    CAS  Google Scholar 

  17. 17

    Yamamoto, A., Cremona, M. L. & Rothman, J. E. Autophagy-mediated clearance of huntingtin aggregates triggered by the insulin-signaling pathway. J. Cell Biol. 172, 719–731 (2006)

    CAS  PubMed  PubMed Central  Google Scholar 

  18. 18

    Yoshikawa, Y. et al. Listeria monocytogenes ActA-mediated escape from autophagic recognition. Nature Cell Biol. 11, 1233–1240 (2009)

    CAS  PubMed  Google Scholar 

  19. 19

    Zhao, Z. et al. Autophagosome-independent essential function for the autophagy protein Atg5 in cellular immunity to intracellular pathogens. Cell Host Microbe 4, 458–469 (2008)

    CAS  PubMed  PubMed Central  Google Scholar 

  20. 20

    Campbell, G. R. & Spector, S. A. Hormonally active vitamin D3 (1α,25-dihydroxycholecalciferol) triggers autophagy in human macrophages that inhibits HIV-1 infection. J. Biol. Chem. 286, 18890–18902 (2011)

    CAS  PubMed  PubMed Central  Google Scholar 

  21. 21

    Campbell, G. R. & Spector, S. A. Vitamin D inhibits human immunodeficiency virus type 1 and Mycobacterium tuberculosis infection in macrophages through the induction of autophagy. PLoS Pathog. 8, e1002689 (2012)

    CAS  PubMed  PubMed Central  Google Scholar 

  22. 22

    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 15, 1101–1111 (2004)

    CAS  PubMed  PubMed Central  Google Scholar 

  23. 23

    Fischer, P. M. The design, synthesis and application of stereochemical and directional peptide isomers: a critical review. Curr. Protein Pept. Sci. 4, 339–356 (2003)

    CAS  PubMed  Google Scholar 

  24. 24

    Couderc, T. et al. A mouse model for Chikungunya: young age and inefficient type-I interferon signaling are risk factors for severe disease. PLoS Pathog. 4, e29 (2008)

    PubMed  PubMed Central  Google Scholar 

  25. 25

    Johnson, R. T. Acute encephalitis. Clin. Infect. Dis. 23, 219–226 (1996)

    CAS  PubMed  Google Scholar 

  26. 26

    Griffin, D. E. Emergence and re-emergence of viral diseases of the central nervous system. Prog. Neurobiol. 91, 95–101 (2010)

    CAS  PubMed  Google Scholar 

  27. 27

    Lauer, P., Chow, M. Y., Loessner, M. J., Portnoy, D. A. & Calendar, R. Construction, characterization, and use of two Listeria monocytogenes site-specific phage integration vectors. J. Bacteriol. 184, 4177–4186 (2002)

    CAS  PubMed  PubMed Central  Google Scholar 

  28. 28

    Sato, M. et al. Multiple oncogenic changes (K-RAS(V12), p53 knockdown, mutant EGFRs, p16 bypass, telomerase) are not sufficient to confer a full malignant phenotype on human bronchial epithelial cells. Cancer Res. 66, 2116–2128 (2006)

    CAS  PubMed  Google Scholar 

  29. 29

    Pattingre, S. et al. Bcl-2 antiapoptotic proteins inhibit Beclin 1-dependent autophagy. Cell 122, 927–939 (2005)

    CAS  PubMed  Google Scholar 

  30. 30

    O’Neill, E. et al. Dynamic evolution of the human immunodeficiency virus type 1 pathogenic factor, Nef. J. Virol. 80, 1311–1320 (2006)

    PubMed  PubMed Central  Google Scholar 

  31. 31

    Kabeya, Y. et al. LC3, a mammalian homologue of yeast Apg8p, is localized in autophagosome membranes after processing. EMBO J. 19, 5720–5728 (2000)

    CAS  PubMed  PubMed Central  Google Scholar 

  32. 32

    Kimura, S., Noda, T. & Yoshimori, T. Dissection of the autophagosome maturation process by a novel reporter protein, tandem fluorescent-tagged LC3. Autophagy 3, 452–460 (2007)

    CAS  PubMed  Google Scholar 

  33. 33

    Thoreen, C. C. et al. An ATP-competitive mammalian target of rapamycin inhibitor reveals rapamycin-resistant functions of mTORC1. J. Biol. Chem. 284, 8023–8032 (2009)

    CAS  PubMed  PubMed Central  Google Scholar 

  34. 34

    Balch, W. E., Dunphy, W. G., Braell, W. A. & Rothman, J. E. Reconstitution of the transport of protein between successive compartments of the Golgi measured by the coupled incorporation of N-acetylglucosamine. Cell 39, 405–416 (1984)

    CAS  PubMed  Google Scholar 

  35. 35

    Vogels, M. W. et al. Quantitative proteomic identification of host factors involved in the Salmonella typhimurium infection cycle. Proteomics 11, 4477–4491 (2011)

    CAS  PubMed  Google Scholar 

  36. 36

    Bréhin, A. C. et al. Production and characterization of mouse monoclonal antibodies reactive to Chikungunya envelope E2 glycoprotein. Virology 371, 185–195 (2008)

    PubMed  Google Scholar 

  37. 37

    Mihalek, I., Res, I. & Lichtarge, O. A family of evolution-entropy hybrid methods for ranking protein residues by importance. J. Mol. Biol. 336, 1265–1282 (2004)

    CAS  PubMed  Google Scholar 

  38. 38

    Bailey, C. K., Andriola, I. F., Kampinga, H. H. & Merry, D. E. Molecular chaperones enhance the degradation of expanded polyglutamine repeat androgen receptor in a cellular model of spinal and bulbar muscular atrophy. Hum. Mol. Genet. 11, 515–523 (2002)

    CAS  PubMed  Google Scholar 

  39. 39

    Taylor, R. M., Hurlbut, H. S., Work, T. H., Kingston, J. R. & Frothingham, T. E. Sindbis virus: a newly recognized arthropod-transmitted virus. Am. J. Trop. Med. Hyg. 4, 844–862 (1955)

    CAS  PubMed  Google Scholar 

  40. 40

    Keller, B. C. et al. Resistance to alpha/beta interferon is a determinant of West Nile virus replication fitness and virulence. J. Virol. 80, 9424–9434 (2006)

    CAS  PubMed  PubMed Central  Google Scholar 

  41. 41

    Weiner, L. P., Cole, G. A. & Nathanson, N. Experimental encephalitis following peripheral inoculation of West Nile virus in mice of different ages. J. Hyg. (Lond.) 68, 435–446 (1970)

    CAS  Google Scholar 

  42. 42

    Schuffenecker, I. et al. Genome microevolution of chikungunya viruses causing the Indian Ocean outbreak. PLoS Med. 3, e263 (2006)

    PubMed  PubMed Central  Google Scholar 

  43. 43

    Gartner, S. et al. The role of mononuclear phagocytes in HTLV-III/LAV infection. Science 233, 215–219 (1986)

    ADS  CAS  PubMed  Google Scholar 

  44. 44

    Qu, X. et al. Promotion of tumorigenesis by heterozygous disruption of the beclin 1 autophagy gene. J. Clin. Invest. 112, 1809–1820 (2003)

    CAS  PubMed  PubMed Central  Google Scholar 

Download references

Acknowledgements

We thank M. Diamond, J. L. Foster, M. Gale, N. Mizushima, D. Sabatini, M. Shiloh and T. Yoshimori for supplying critical reagents; and H. Ball, A. Bugde and E.-L. Eskelinen for assistance with peptide synthesis, infrared imaging and EM interpretation, respectively. This work was supported by NIH grants U54AI057156 (B.L.), K08 AI099150 (R.S.), ROI NS077874 (S.A.S), RO1 GM094575 (N.V.G.), ROI GM066099 (O.L.), ROI GM079656 (O.L.), ROI NS063973 (A.Y.), ROI NS050199 (A.Y.), U54AI057160 (H.W.V., D.J.L.), ROI DK083756 (R.X.), ROI DK086502 (R.X.), and T32 GM008297 (C.H.); NSF CCF-0905536 (O.L.); an NWO-ALW Open Program Grant 817.02.023 (J.B.H.); Cancer Research UK (S.A.T.); and a Welch Foundation Award I-15-5 (N.V.G.).

Author information

Affiliations

Authors

Contributions

B.L., S.S.-K. and O.L. generated the original hypothesis. S.S.-K., R.S., M.L., G.R.C., Z.Z., Q.S., K.P., D.M., C.H., R.E., D.K. and D.V.K. performed experiments. L.K., A.D.W., R.X., O.L. and N.V.G. performed bioinformatics analyses. B.L., S.S.-K., R.S., M.L., L.K., H.W.V., J.B.H., S.A.T., R.X., D.J.L., A.Y., O.L., N.V.G., S.A.S. and D.V.K. provided intellectual contributions throughout the project. B.L., S.S.-K. and R.S. took primary responsibility for writing the manuscript. All authors edited the manuscript.

Corresponding author

Correspondence to Beth Levine.

Ethics declarations

Competing interests

The authors declare no competing financial interests.

Supplementary information

Supplementary Information

This file contains Supplementary Figures 1-28 and Supplementary Table 1. This file was replaced on 13 February 2013 as the original had corrupted. (PDF 2373 kb)

PowerPoint slides

Rights and permissions

Reprints and Permissions

About this article

Cite this article

Shoji-Kawata, S., Sumpter, R., Leveno, M. et al. Identification of a candidate therapeutic autophagy-inducing peptide. Nature 494, 201–206 (2013). https://doi.org/10.1038/nature11866

Download citation

Further reading

Comments

By submitting a comment you agree to abide by our Terms and Community Guidelines. If you find something abusive or that does not comply with our terms or guidelines please flag it as inappropriate.

Search

Nature Briefing

Sign up for the Nature Briefing newsletter — what matters in science, free to your inbox daily.

Get the most important science stories of the day, free in your inbox. Sign up for Nature Briefing