Abstract
Solar ultraviolet radiations induced DNA damages in human skin cells with cyclobutane pyrimidine dimers (CPD) and (6–4) photoproducts (6–4PPs) as the most frequent lesions. CPDs are repaired much slower than 6–4PPs by the nucleotide excision repair pathway, which are thus the major lesions that interfere with key cellular processes and give rise to gene mutations, possibly resulting in skin cancer. In prokaryotes and multicellular eukaryotes other than placental mammals, CPDs can be rapidly repaired by CPD photolyases in one simple enzymatic reaction using the energy of blue light. In this study, we aim to construct recombinant CPD photolyases that can autonomously enter human cell nuclei to fix UV-induced CPDs. A fly cell penetration peptide and a viral nucleus localization signal peptide were recombined with a fungal CPD photolyase to construct a recombinant protein. This engineered CPD photolyase autonomously crosses cytoplasm and nuclear membrane of human cell nuclei, which then efficiently photo-repairs UV-induced CPD lesions in the genomic DNA. This further protects the cells by increasing SOD activity, and decreasing cellular ROSs, malondialdehyde and apoptosis.
Similar content being viewed by others
Data availability
All relevant data supporting the findings of the study are available in this article and its Supplementary Information files, or from the corresponding author upon request.
References
Abri Aghdam M, Bagheri R, Mosafer J, Baradaran B, Hashemzaei M, Baghbanzadeh A, de la Guardia M, Mokhtarzadeh A (2019) Recent advances on thermosensitive and pH-sensitive liposomes employed in controlled release. J Control Release 315:1–22. https://doi.org/10.1016/j.jconrel.2019.09.018
Banskota S, Raguram A, Suh S, Du SW, Davis JR, Choi EH, Wang X, Nielsen SC, Newby GA, Randolph PB, Osborn MJ, Musunuru K, Palczewski K, Liu DR (2022) Engineered virus-like particles for efficient in vivo delivery of therapeutic proteins. Cell 185(2):250-265.e16. https://doi.org/10.1016/j.cell.2021.12.021
Bolhassani A, Jafarzade BS, Mardani G (2017) In vitro and in vivo delivery of therapeutic proteins using cell penetrating peptides. Peptides 87:50–63. https://doi.org/10.1016/j.peptides.2016.11.011
Boros G, Miko E, Muramatsu H, Weissman D, Emri E, Rozsa D, Nagy G, Juhasz A, Juhasz I, van der Horst GT, Horkay I, Remenyik E, Kariko K, Emri G (2013) Transfection of pseudouridine-modified mRNA encoding cpd-photolyase leads to repair of dna damage in human keratinocytes: a new approach with future therapeutic potential. J Photoch Photobio B: Biology 129:93–99. https://doi.org/10.1016/j.jphotobiol.2013.09.010
Chiganças V, Miyaji EN, Muotri AR, de Fátima Jacysyn J, Amarante-Mendes GP, Yasui A, Menck CF (2000) Photorepair prevents ultraviolet-induced apoptosis in human cells expressing the marsupial photolyase gene. Cancer Res, 60(9), 2458–2463. https://pubmed.ncbi.nlm.nih.gov/10811124/. Accessed Jan 06 2014
Del Rio D, Stewart AJ, Pellegrini N (2005) A review of recent studies on malondialdehyde as toxic molecule and biological marker of oxidative stress. Nutr Metab Cardiovas: NMCD 15(4):316–328. https://doi.org/10.1016/j.numecd.2005.05.003
Derossi D, Joliot AH, Chassaing G, Prochiantz A (1994) The third helix of the Antennapedia homeodomain translocates through biological membranes. J Biol Chem 269(14):10444–10450
Fang W, St. Leger RJ, (2012) Enhanced UV Resistance and Improved Killing of Malaria Mosquitoes by Photolyase Transgenic Entomopathogenic Fungi. PLoS ONE 7(8):e43069. https://doi.org/10.2217/14796694.2.2.191
Garinis GA, Jans J, van der Horst GT (2006) Photolyases: capturing the light to battle skin cancer. Future Oncol (london, England) 2(2):191–199. https://doi.org/10.2217/14796694.2.2.191
Guintini L, Charton R, Peyresaubes F, Thoma F, Conconi A (2015) Nucleosome positioning, nucleotide excision repair and photoreactivation in Saccharomyces cerevisiae. DNA Repair 36:98–104. https://doi.org/10.1016/j.dnarep.2015.09.012
Leccia MT, Lebbe C, Claudel JP, Narda M, Basset-Seguin N (2019) New Vision in Photoprotection and Photorepair. Dermatology Ther 9(1):103–115. https://doi.org/10.1007/s13555-019-0282-5
Lee TH, Kang TH (2019) DNA Oxidation and Excision Repair Pathways. Int J Mol Sci 20(23):6092. https://doi.org/10.3390/ijms20236092
Liu W, Ye A, Han F, Ha J (2019) Advances and challenges in liposome digestion: Surface interaction, biological fate, and GIT modeling. Adv Colloid Interfac 263:52–67. https://doi.org/10.1016/j.cis.2018.11.007
Marteijn JA, Lans H, Vermeulen W, Hoeijmakers JH (2014) Understanding nucleotide excision repair and its roles in cancer and ageing. Nat Rev Mol Cell Biol 15:465–481. https://doi.org/10.1038/nrm382
Sancar A (2003) Structure and function of DNA photolyase and cryptochrome blue-light photoreceptors. Chem Rev 103(6):2203–2237. https://doi.org/10.1021/cr0204348
Sancar A (2016) Mechanismen der DNA-Reparatur durch Photolyasen und Exzisionsnukleasen (Nobel-Aufsatz). Angew Chem Int Edit 128(30):8643–8670. https://doi.org/10.1002/anie.201601524
Schul W, Jans J, Rijksen YM, Klemann KH, Eker AP, de Wit J, Nikaido O, Nakajima S, Yasui A, Hoeijmakers JH, van der Horst GT (2002) Enhanced repair of cyclobutane pyrimidine dimers and improved UV resistance in photolyase transgenic mice. EMBO J 21(17):4719–4729. https://doi.org/10.1093/emboj/cdf456
Stege H, Roza L, Vink AA, Grewe M, Ruzicka T, Grether-Beck S, Krutmann J (2000) Enzyme plus light therapy to repair DNA damage in ultraviolet -B-irradiated human skin. P Natl Acad Sci USA 97(4):1790–1795. https://doi.org/10.1073/pnas.030528897
Verma R, Pal S, Kumar J, Haldar C (2017) Ultraviolet a (UV-A) radiation-induced damage in the skin and vital organs of albino rat: an indirect correlation with melatonin. J Endo Repr 2:59–66. https://doi.org/10.18311/jer/2017/21626
Yasui A, Eker AP, Yasuhira S, Yajima H, Kobayashi T, Takao M, Oikawa A (1994) A new class of DNA photolyases present in various organisms including aplacental mammals. EMBO J 13(24):6143–6151. https://doi.org/10.1002/j.1460-2075.1994.tb06961.x
Yi C, He C (2013) DNA repair by reversal of DNA damage. CSH Prspect Biol 5(1):a012575. https://doi.org/10.1101/cshperspect.a012575
You YH, Lee DH, Yoon JH, Nakajima S, Yasui A, Pfeifer GP (2001) Cyclobutane pyrimidine dimers are responsible for the vast majority of mutations induced by UVB irradiation in mammalian cells. J Biol Chem 276(48):44688–44694. https://doi.org/10.1074/jbc.M1FIG07696200
Zeng G, Chen X, Zhang X, Zhang Q, Xu C, Mi W, Guo N, Zhao H, You Y, Dryburgh FJ, Bidochka MJ, St Leger RJ, Zhang L, Fang W (2017) Genome-wide identification of pathogenicity, conidiation and colony sectorization genes in Metarhizium robertsii. Environ Microbiol 19(10):3896–3908. https://doi.org/10.1111/1462-2920.13777
Zhang M, Wang L, Zhong D (2017) Photolyase: Dynamics and electron-transfer mechanisms of DNA repair. Arch Biochem Biophys 632:158–174. https://doi.org/10.1016/j.abb.2017.08.007
Zhang Q, Chen X, Xu C, Zhao H, Zhang X, Zeng G, Qian Y, Liu R, Guo N, Mi W, Meng Y, Leger R, Fang W (2019) Horizontal gene transfer allowed the emergence of broad host range entomopathogens. P Natl Acad Sci USA 116(16):7982–7989. https://doi.org/10.1073/pnas.1816430116
Funding
This work was supported by National Natural Science Foundation of China Grants (32172470).
Author information
Authors and Affiliations
Contributions
WF designed research. YB performed research. YB analyzed data; YB and WF wrote the paper. All authors read and approved the final manuscript.
Corresponding author
Ethics declarations
Conflict of interest
The authors declare no competing interests.
Ethics approval
This article does not contain any studies with human participants or animals performed by any of the authors.
Additional information
Publisher's Note
Springer Nature remains neutral with regard to jurisdictional claims in published maps and institutional affiliations.
Supplementary Information
Below is the link to the electronic supplementary material.
Rights and permissions
Springer Nature or its licensor (e.g. a society or other partner) holds exclusive rights to this article under a publishing agreement with the author(s) or other rightsholder(s); author self-archiving of the accepted manuscript version of this article is solely governed by the terms of such publishing agreement and applicable law.
About this article
Cite this article
Bao, Y., Fang, W. A recombinant fungal photolyase autonomously enters human cell nuclei to fix UV-induced DNA lesions. Biotechnol Lett 46, 459–467 (2024). https://doi.org/10.1007/s10529-024-03474-3
Received:
Revised:
Accepted:
Published:
Issue Date:
DOI: https://doi.org/10.1007/s10529-024-03474-3