Journal Home > Volume 13 , Issue 12
Nano Research 2020, 13(12): 3253-3260 https://doi.org/10.1007/s12274-020-2998-z
Research Article | Issue | Published: 14 August 2020

Spatiotemporal regulation of ubiquitin-mediated protein degradation via upconversion optogenetic nanosystem

Show Author's Information Yafeng Hao1,§,  Taofeng Du2,§,  Gaoju Pang1,  Jiahua Li1,  Huizhuo Pan1,  Yingying Zhang1,  Lizhen Wang2,  Jin Chang1,  En-min Zhou2( ),  Hanjie Wang1( )
School of Life Sciences, Tianjin University, Tianjin 300072, China
Department of preventive Veterinary Medicine, College of Veterinary Medicine, Northwest A&F University, Yangling 712100, China

§ Yafeng Hao and Taofeng Du contributed equally to this work.

Keywords:
optogenetics, protein degradation, ubiquitin-proteasome system, upconversion materials, the nonstructural protein9 (NSP9), porcine reproductive and respiratory syndrome virus (PRRSV)
Topics:
ProteinsViruses
Cite this article:
Hao Y, Du T, Pang G, et al. Spatiotemporal regulation of ubiquitin-mediated protein degradation via upconversion optogenetic nanosystem. Nano Research, 2020, 13(12): 3253-3260. https://doi.org/10.1007/s12274-020-2998-z
Download citation
EndNote(RIS)
BibTeX

541

Views

18

Downloads

Citations

4

Crossref

N/A

WoS

3

Scopus

0

CSCD

Abstract Full text Electronic supplementary material About this article

Protein degradation technology, which is one of the most direct and effective ways to regulate the life activities of cells, is expected to be applied to the treatment of various diseases. However, current protein degradation technologies such as some small-molecule degraders which are unable to achieve spatiotemporal regulation, making them difficult to transform into clinical applications. In this article, an upconversion optogenetic nanosystem was designed to attain accurate regulation of protein degradation. This system worked via two interconnected parts: 1) the host cell expressed light-sensitive protein that could trigger the ubiquitin-proteasome pathway upon blue-light exposure; 2) the light regulated light-sensitive protein by changing light conditions to achieve regulation of protein degradation. Experimental results based on model protein (Green Fluorescent Protein, GFP) validated that this system could fulfill protein degradation both in vitro (both Hela and 293T cells) and in vivo (by upconversion optogenetic nanosystem), and further demonstrated that we could reach spatiotemporal regulation by changing the illumination time (0-25 h) and the illumination frequency (the illuminating frequency of 0-30 s every 1 min). We further took another functional protein (The Nonstructural Protein 9, NSP9) into experiment. Results confirmed that the proliferation of porcine reproductive and respiratory syndrome virus (PRRSV) was inhibited by degrading the NSP9 in this light-induced system, and PRRSV proliferation was affected by different light conditions (illumination time varies from 0-24 h). We expected this system could provide new perspectives into spatiotemporal regulation of protein degradation and help realize the clinical application transformation for treating diseases of protein degradation technology.


menu
Abstract
Full text
Outline
Electronic supplementary material
About this article

Spatiotemporal regulation of ubiquitin-mediated protein degradation via upconversion optogenetic nanosystem

Show Author's information Yafeng Hao1,§, Taofeng Du2,§, Gaoju Pang1, Jiahua Li1, Huizhuo Pan1, Yingying Zhang1, Lizhen Wang2, Jin Chang1, En-min Zhou2( ), Hanjie Wang1( )
School of Life Sciences, Tianjin University, Tianjin 300072, China
Department of preventive Veterinary Medicine, College of Veterinary Medicine, Northwest A&F University, Yangling 712100, China

§ Yafeng Hao and Taofeng Du contributed equally to this work.

Abstract

Protein degradation technology, which is one of the most direct and effective ways to regulate the life activities of cells, is expected to be applied to the treatment of various diseases. However, current protein degradation technologies such as some small-molecule degraders which are unable to achieve spatiotemporal regulation, making them difficult to transform into clinical applications. In this article, an upconversion optogenetic nanosystem was designed to attain accurate regulation of protein degradation. This system worked via two interconnected parts: 1) the host cell expressed light-sensitive protein that could trigger the ubiquitin-proteasome pathway upon blue-light exposure; 2) the light regulated light-sensitive protein by changing light conditions to achieve regulation of protein degradation. Experimental results based on model protein (Green Fluorescent Protein, GFP) validated that this system could fulfill protein degradation both in vitro (both Hela and 293T cells) and in vivo (by upconversion optogenetic nanosystem), and further demonstrated that we could reach spatiotemporal regulation by changing the illumination time (0-25 h) and the illumination frequency (the illuminating frequency of 0-30 s every 1 min). We further took another functional protein (The Nonstructural Protein 9, NSP9) into experiment. Results confirmed that the proliferation of porcine reproductive and respiratory syndrome virus (PRRSV) was inhibited by degrading the NSP9 in this light-induced system, and PRRSV proliferation was affected by different light conditions (illumination time varies from 0-24 h). We expected this system could provide new perspectives into spatiotemporal regulation of protein degradation and help realize the clinical application transformation for treating diseases of protein degradation technology.

Keywords: optogenetics, protein degradation, ubiquitin-proteasome system, upconversion materials, the nonstructural protein9 (NSP9), porcine reproductive and respiratory syndrome virus (PRRSV)

References(35)

[1]
N. K. Verma,; A. Gadi,; G. Maurizi,; U. B. Roy,; A. Mansukhani,; C. Basilico, Myeloid zinc finger 1 and GA binding protein co-operate with Sox2 in regulating the expression of yes-associated protein 1 in cancer cells. Stem Cells 2017, 35, 2340-2350.
Google Scholar
[2]
X. G. Huang,; G. H. Zhou,; W. Y. Wu,; Y. J. Duan,; G. E. Ma,; J. Y. Song,; R. Xiao,; L. Vandenberghe,; F. Zhang,; P. A. D’Amore, et al. Genome editing abrogates angiogenesis in vivo. Nat. Commun. 2017, 8, 112.
Google Scholar
[3]
L. A. Becker,; B. Huang,; G. Bieri,; R. Ma,; D. A. Knowles,; P. Jafar-Nejad,; J. Messing,; H. J. Kim,; A. Soriano,; G. Auburger, et al. Therapeutic reduction of ataxin-2 extends lifespan and reduces pathology in TDP-43 mice. Nature 2017, 544, 367-371.
Google Scholar
[4]
Y. L. Wu,; H. C. Zhou,; G. Q. Chen, Molecular mechanisms of leukemia-associated protein degradation. Front. Med. China 2010, 4, 363-70.
Google Scholar
[5]
E. W. Lee,; J. H. Kim,; Y. H. Ahn,; J. Seo,; A. Ko,; M. Jeong,; S. J. Kim,; J. Y. Ro,; K. M. Park,; H. W. Lee, et al. Ubiquitination and degradation of the FADD adaptor protein regulate death receptor-mediated apoptosis and necroptosis. Nat. Commun. 2012, 3, 978.
Google Scholar
[6]
J. M. Heo,; N. Livnat-Levanon,; E. B. Taylor,; K. T. Jones,; N. Dephoure,; J. Ring,; J. X. Xie,; J. L. Brodsky,; F. Madeo,; S. P. Gygi, et al. A stress-responsive system for mitochondrial protein degradation. Mol. Cell 2010, 40, 465-480.
Google Scholar
[7]
D. M. Koepp, Cell cycle regulation by protein degradation. In: Cell Cycle Control. E. Noguchi,; M. Gadaleta,, Eds.; Humana Press: New York, 2014; pp 61-73.
[8]
D. Clift,; W. A. McEwan,; L. I. Labzin,; V. Konieczny,; B. Mogessie,; L. C. James,; M. Schuh, A method for the acute and rapid degradation of endogenous proteins. Cell 2017, 171, 1692-1706.e18.
Google Scholar
[9]
A. Aguzzi,; T. O’Connor, Protein aggregation diseases: Pathogenicity and therapeutic perspectives. Nat. Rev. Drug Discov. 2010, 9, 237-248.
Google Scholar
[10]
M. Pettersson,; C. M. Crews, Proteolysis targeting chimeras (PROTACs)-past, present and future. Drug Discov. Today Technol. 2019, 31, 15-27.
Google Scholar
[11]
P. P. Chamberlain,; L. G. Hamann, Development of targeted protein degradation therapeutics. Nat. Chem. Biol. 2019, 15, 937-944.
Google Scholar
[12]
P. Ottis,; C. M. Crews, Proteolysis-targeting chimeras: Induced protein degradation as a therapeutic strategy. ACS Chem. Biol. 2017, 12, 892-898.
Google Scholar
[13]
E. M. Aro,; R. Kettunen,; E. Tyystjärvi, ATP and light regulate D1 protein modification and degradation role of D1* in photoinhibition. FEBS Lett. 1992, 297, 29-33.
Google Scholar
[14]
J. C. Groppe, Induced degradation of protein kinases by bifunctional small molecules: A next-generation strategy. Expert Opin. Drug Discov. 2019, 14, 1237-1253.
Google Scholar
[15]
M. A. Eldeeb,; R. Siva-Piragasam,; M. A. Ragheb,; M. Esmaili,; M. Salla,; R. P. Fahlman, A molecular toolbox for studying protein degradation in mammalian cells. J. Neurochem. 2019, 151, 520-533.
Google Scholar
[16]
L. Fenno,; O. Yizhar,; K. Deisseroth, The development and application of optogenetics. Annu. Rev. Neurosci. 2011, 34, 389-412.
Google Scholar
[17]
J. Duebel,; K. Marazova,; J. A. Sahel, Optogenetics. Curr. Opin. Ophthalmol. 2015, 26, 226-232.
Google Scholar
[18]
H. Y. Jia,; L. Kai,; M. Heymann,; D. A. Garcia-Soriano,; T. Härtel,; P. Schwille, Light-induced printing of protein structures on membranes in vitro. Nano Lett. 2018, 18, 7133-7140.
Google Scholar
[19]
P. Jayaraman,; J. W. Yeoh,; S. Jayaraman,; A. Y. Teh,; J. Y. Zhang,; C. L. Poh, Cell-free optogenetic gene expression system. ACS Synth. Biol. 2018, 7, 986-994.
Google Scholar
[20]
A. Galvan,; W. R. Stauffer,; L. Acker,; Y. El-Shamayleh,; K. I. Inoue,; S. Ohayon,; M. C. Schmid, Nonhuman primate optogenetics: Recent advances and future directions. J. Neurosci. 2017, 37, 10894-10903.
Google Scholar
[21]
M. Yamada,; Y. Suzuki,; S. C. Nagasaki,; H. Okuno,; I. Imayoshi, Light control of the tet gene expression system in mammalian cells. Cell Rep. 2018, 25, 487-500.E6.
Google Scholar
[22]
P. Tan,; L. He,; G. Han,; Y. B. Zhou, Optogenetic immunomodulation: Shedding light on antitumor immunity. Trends Biotechnol. 2017, 35, 215-226.
Google Scholar
[23]
B. Zheng,; H. J. Wang,; H. Z. Pan,; C. Liang,; W. Y. Ji,; L. Zhao,; H. B. Chen,; X. Q. Gong,; X. L. Wu,; J. Chang, Near-infrared light triggered upconversion optogenetic nanosystem for cancer therapy. ACS Nano 2017, 11, 11898-11907.
Google Scholar
[24]
H. Z. Pan,; H. J. Wang,; J. X. Yu,; X. Huang,; Y. F. Hao,; C. N. Zhang,; W. Y. Ji,; M. Y. Yang,; X. Q. Gong,; X. L. Wu, et al. Near-infrared light remotely up-regulate autophagy with spatiotemporal precision via upconversion optogenetic nanosystem. Biomaterials 2019, 199, 22-31.
Google Scholar
[25]
S. Chen,; A. Z. Weitemier,; X. Zeng,; L. M. He,; X. Y. Wang,; Y. Q. Tao,; A. J. Y. Huang,; Y. Hashimotodani,; M. Kano,; H. Iwasaki, et al. Near-infrared deep brain stimulation via upconversion nanoparticle-mediated optogenetics. Science 2018, 359, 679-684.
Google Scholar
[26]
X. Wang,; X. J. Chen,; Y. Yang, Spatiotemporal control of gene expression by a light-switchable transgene system. Nat. Methods 2012, 9, 266-269.
Google Scholar
[27]
D. A. Rhodes,; D. A. Isenberg, TRIM21 and the function of antibodies inside cells. Trends Immunol. 2017, 38, 916-926.
Google Scholar
[28]
D. L. Mallery,; W. A. McEwan,; S. R. Bidgood,; G. J. Towers,; C. M. Johnson,; L. C. James, Antibodies mediate intracellular immunity through tripartite motif-containing 21 (TRIM21). Proc. Natl. Acad. Sci. USA 2010, 107, 19985-19990.
Google Scholar
[29]
S. W. De Taeye,; T. Rispens,; G. Vidarsson, The ligands for human IgG and their effector functions. Antibodies 2019, 8, 30.
Google Scholar
[30]
Y. Fang,; E. J. Snijder, The PRRSV replicase: Exploring the multifunctionality of an intriguing set of nonstructural proteins. Virus Res. 2010, 154, 61-76.
Google Scholar
[31]
L. Liu,; J. Tian,; H. Nan,; M. M. Tian,; Y. Li,; X. D. Xu,; B. C. Huang,; E. M. Zhou,; J. A. Hiscox,; H. Y. Chen, Porcine reproductive and respiratory syndrome virus nucleocapsid protein interacts with Nsp9 and cellular DHX9 to regulate viral RNA synthesis. J. Virol. 2016, 90, 5384-5398.
Google Scholar
[32]
L. Z. Wang,; L. Zhang,; B. C. Huang,; K. K. Li,; G. P. Hou,; Q. Zhao,; C. Y. Wu,; Y. C. Nan,; T. F. Du,; Y. Mu, et al. A nanobody targeting viral nonstructural protein 9 inhibits porcine reproductive and respiratory syndrome virus replication. J. Virol. 2019, 93, e01888-18 .
Google Scholar
[33]
D. Virology. Normile, China, vietnam grapple with ‘rapidly evolving’ pig virus. Science 2007, 317, 1017.
Google Scholar
[34]
M. P. Murtaugh,; M. Genzow, Immunological solutions for treatment and prevention of porcine reproductive and respiratory syndrome (PRRS). Vaccine 2011, 29, 8192-8204.
Google Scholar
[35]
H. L. Liu,; Y. Wang,; H. Duan,; A. Zhang,; C. Liang,; J. M. Gao,; C. Zhang,; B. C. Huang,; Q. Y. Li,; N. Li, et al. An intracellularly expressed Nsp9-specific nanobody in MARC-145 cells inhibits porcine reproductive and respiratory syndrome virus replication. Vet. Microbiol. 2015, 181, 252-260.
Google Scholar
File
12274_2020_2998_MOESM1_ESM.pdf (2.1 MB)
Publication history
Copyright
Acknowledgements

Publication history

Received: 28 January 2020
Revised: 17 July 2020
Accepted: 20 July 2020
Published: 14 August 2020
Issue date: December 2020

Copyright

© Tsinghua University Press and Springer-Verlag GmbH Germany, part of Springer Nature

Acknowledgements

This work was sponsored by the National Key Research and Development Program of China (Nos. 2019YFA0906500 and 2017YFA0205104), the National Natural Science Foundation of China (Nos. 31971300, 817719709, 51873150 and 51573128), Tianjin Natural Science Foundation (No. 19JCYBJC28800) and Young Elite Scientists Sponsorship Program by Tianjin.

Return