Initiation of Repair of DNA−Polypeptide Cross-Links by the UvrABC Nuclease† (original) (raw)

ArticleFebruary 1, 2005

Click to copy article linkArticle link copied!

Biochemistry

Cite this: Biochemistry 2005, 44, 8

Click to copy citationCitation copied!

Published February 1, 2005

research-article

Copyright © 2005 American Chemical Society

Abstract

Click to copy section linkSection link copied!

Abstract Image

Although the biochemical pathways that repair DNA−protein cross-links have not been clearly elucidated, it has been proposed that the partial proteolysis of cross-linked proteins into smaller oligopeptides constitutes an initial step in removal of these lesions by nucleotide excision repair (NER). To test the validity of this repair model, several site-specific DNA−peptide and DNA−protein cross-links were engineered via linkage at (1) an acrolein-derived γ-hydroxypropanodeoxyguanosine adduct and (2) an apurinic/apyrimidinic site, and the initiation of repair was examined in vitro using recombinant proteins UvrA and UvrB from Bacillus caldotenax and UvrC from Thermotoga maritima. The polypeptides cross-linked to DNA were Lys-Trp-Lys-Lys, Lys-Phe-His-Glu-Lys-His-His-Ser-His-Arg-Gly-Tyr, and the 16 kDa protein, T4 pyrimidine dimer glycosylase/apurinic/apyrimidinic site lyase. For the substrates examined, DNA incision required the coordinated action of all three proteins and occurred at the eighth phosphodiester bond 5' to the lesion. The incision rates for DNA−peptide cross-links were comparable to or greater than that measured on fluorescein-adducted DNA, an excellent substrate for UvrABC. Incision rates were dependent on both the site of covalent attachment on the DNA and the size of the bound peptide. Importantly, incision of a DNA−protein cross-link occurred at a rate approximately 3.5−8-fold slower than the rates observed for DNA−peptide cross-links. Thus, direct evidence has been obtained indicating that (1) DNA−peptide cross-links can be efficiently incised by the NER proteins and (2) DNA−peptide cross-links are preferable substrates for this system relative to DNA−protein cross-links. These data suggest that proteolytic degradation of DNA−protein cross-links may be an important processing step in facilitating NER.

ACS Publications

Copyright © 2005 American Chemical Society

Cited By

Click to copy section linkSection link copied!

This article is cited by 63 publications.

  1. Cameron Bryan, Joel Cepeda, Bingru Li, Kun Yang. DNA–Protein Cross-Links Derived from Abasic DNA Lesions: Recent Progress and Future Directions. Chemical Research in Toxicology 2025, 38 (6) , 997-1005. https://doi.org/10.1021/acs.chemrestox.5c00125
  2. Tingyu Wen, Shubo Zhao, Julian Stingele, Jean-Luc Ravanat, Marc M. Greenberg. Quantification of Intracellular DNA–Protein Cross-Links with N7-Methyl-2′-Deoxyguanosine and Their Contribution to Cytotoxicity. Chemical Research in Toxicology 2024, 37 (5) , 814-823. https://doi.org/10.1021/acs.chemrestox.4c00076
  3. Rebekah M. B. Silva, Michael A. Grodick, Jacqueline K. Barton. UvrC Coordinates an O2-Sensitive [4Fe4S] Cofactor. Journal of the American Chemical Society 2020, 142 (25) , 10964-10977. https://doi.org/10.1021/jacs.0c01671
  4. Natalia Y. Tretyakova, Arnold Groehler, IV, and Shaofei Ji . DNA–Protein Cross-Links: Formation, Structural Identities, and Biological Outcomes. Accounts of Chemical Research 2015, 48 (6) , 1631-1644. https://doi.org/10.1021/acs.accounts.5b00056
  5. Jung Eun Yeo, Susith Wickramaratne, Santoshkumar Khatwani, Yen-Chih Wang, Jeffrey Vervacke, Mark D. Distefano, and Natalia Y. Tretyakova . Synthesis of Site-Specific DNA–Protein Conjugates and Their Effects on DNA Replication. ACS Chemical Biology 2014, 9 (8) , 1860-1868. https://doi.org/10.1021/cb5001795
  6. Susith Wickramaratne, Shivam Mukherjee, Peter W. Villalta, Orlando D. Schärer, and Natalia Y. Tretyakova . Synthesis of Sequence-Specific DNA–Protein Conjugates via a Reductive Amination Strategy. Bioconjugate Chemistry 2013, 24 (9) , 1496-1506. https://doi.org/10.1021/bc400018u
  7. Paige L. McKibbin, Aaron M. Fleming, Mohammad Atif Towheed, Bennett Van Houten, Cynthia J. Burrows, and Sheila S. David . Repair of Hydantoin Lesions and Their Amine Adducts in DNA by Base and Nucleotide Excision Repair. Journal of the American Chemical Society 2013, 135 (37) , 13851-13861. https://doi.org/10.1021/ja4059469
  8. Hai Huang, Hao Wang, Markus W. Voehler, Albena Kozekova, Carmelo J. Rizzo, Amanda K. McCullough, R. Stephen Lloyd, and Michael P. Stone . γ-Hydroxy-1,N2-propano-2′-deoxyguanosine DNA Adduct Conjugates the N-Terminal Amine of the KWKK Peptide via a Carbinolamine Linkage. Chemical Research in Toxicology 2011, 24 (7) , 1123-1133. https://doi.org/10.1021/tx200113n
  9. Hai Huang, Ivan D. Kozekov, Albena Kozekova, Carmelo J. Rizzo, Amanda K. McCullough, R. Stephen Lloyd and Michael P. Stone . Minor Groove Orientation of the KWKK Peptide Tethered via the N-Terminal Amine to the Acrolein-Derived 1,N2-γ-Hydroxypropanodeoxyguanosine Lesion with a Trimethylene Linkage,. Biochemistry 2010, 49 (29) , 6155-6164. https://doi.org/10.1021/bi100364f
  10. Elisabeth M. Loecken, Surendra Dasari, Salisha Hill, David L. Tabb and F. Peter Guengerich. The bis-Electrophile Diepoxybutane Cross-Links DNA to Human Histones but Does Not Result in Enhanced Mutagenesis in Recombinant Systems. Chemical Research in Toxicology 2009, 22 (6) , 1069-1076. https://doi.org/10.1021/tx900037u
  11. Irina G. Minko, Ivan D. Kozekov, Thomas M. Harris, Carmelo J. Rizzo, R. Stephen Lloyd and Michael P. Stone. Chemistry and Biology of DNA Containing 1,N2-Deoxyguanosine Adducts of the α,β-Unsaturated Aldehydes Acrolein, Crotonaldehyde, and 4-Hydroxynonenal. Chemical Research in Toxicology 2009, 22 (5) , 759-778. https://doi.org/10.1021/tx9000489
  12. Irina G. Minko, Kinrin Yamanaka, Ivan D. Kozekov, Albena Kozekova, Chiara Indiani, Michael E. O’Donnell, Qingfei Jiang, Myron F. Goodman, Carmelo J. Rizzo and R. Stephen Lloyd . Replication Bypass of the Acrolein-Mediated Deoxyguanine DNA-Peptide Cross-Links by DNA Polymerases of the DinB Family. Chemical Research in Toxicology 2008, 21 (10) , 1983-1990. https://doi.org/10.1021/tx800174a
  13. Aley G. Kalapila, Natalia A. Loktionova and Anthony E. Pegg. Alkyltransferase-Mediated Toxicity of 1,3-Butadiene Diepoxide. Chemical Research in Toxicology 2008, 21 (9) , 1851-1861. https://doi.org/10.1021/tx800178t
  14. Elisabeth M. Loecken and F. Peter Guengerich. Reactions of Glyceraldehyde 3-Phosphate Dehydrogenase Sulfhydryl Groups with Bis-Electrophiles Produce DNA–Protein Cross-Links but Not Mutations. Chemical Research in Toxicology 2008, 21 (2) , 453-458. https://doi.org/10.1021/tx7003618
  15. Xiaoyun Xu,, James G. Muller,, Yu Ye, and, Cynthia J. Burrows. DNA−Protein Cross-links between Guanine and Lysine Depend on the Mechanism of Oxidation for Formation of C5 Vs C8 Guanosine Adducts. Journal of the American Chemical Society 2008, 130 (2) , 703-709. https://doi.org/10.1021/ja077102a
  16. James J. Truglio,, Deborah L. Croteau,, Bennett Van Houten, and, Caroline Kisker. Prokaryotic Nucleotide Excision Repair: The UvrABC System. Chemical Reviews 2006, 106 (2) , 233-252. https://doi.org/10.1021/cr040471u
  17. Patrick J. O'Brien. Catalytic Promiscuity and the Divergent Evolution of DNA Repair Enzymes. Chemical Reviews 2006, 106 (2) , 720-752. https://doi.org/10.1021/cr040481v
  18. Anna V. Yudkina, Dmitry O. Zharkov. The hidden elephant: Modified abasic sites and their consequences. DNA Repair 2025, 148 , 103823. https://doi.org/10.1016/j.dnarep.2025.103823
  19. Anna V Yudkina, Nikita A Bulgakov, Daria V Kim, Svetlana V Baranova, Alexander A Ishchenko, Murat K Saparbaev, Vladimir V Koval, Dmitry O Zharkov. Abasic site–peptide cross-links are blocking lesions repaired by AP endonucleases. Nucleic Acids Research 2023, 51 (12) , 6321-6336. https://doi.org/10.1093/nar/gkad423
  20. Anna V. Yudkina, Alexander E. Barmatov, Nikita A. Bulgakov, Elizaveta O. Boldinova, Evgeniy S. Shilkin, Alena V. Makarova, Dmitry O. Zharkov. Bypass of Abasic Site–Peptide Cross-Links by Human Repair and Translesion DNA Polymerases. International Journal of Molecular Sciences 2023, 24 (13) , 10877. https://doi.org/10.3390/ijms241310877
  21. Cameron Bryan, Jennifer Le, Xiaoying Wei, Kun Yang. Saccharomyces cerevisiae apurinic/apyrimidinic endonuclease 1 repairs abasic site-mediated DNA-peptide/protein cross-links. DNA Repair 2023, 126 , 103501. https://doi.org/10.1016/j.dnarep.2023.103501
  22. Cameron Bryan, Xiaoying Wei, Zhishuo Wang, Kun Yang. In vitro eradication of abasic site-mediated DNA–peptide/protein cross-links by Escherichia coli long-patch base excision repair. Journal of Biological Chemistry 2022, 298 (7) , 102055. https://doi.org/10.1016/j.jbc.2022.102055
  23. Megan Perry, Gargi Ghosal. Mechanisms and Regulation of DNA-Protein Crosslink Repair During DNA Replication by SPRTN Protease. Frontiers in Molecular Biosciences 2022, 9 https://doi.org/10.3389/fmolb.2022.916697
  24. Suresh S. Pujari, Mingxuan Wu, Jenna Thomforde, Zhipeng A. Wang, Christopher Chao, Noelle M. Olson, Luke Erber, William C. K. Pomerantz, Philip Cole, Natalia Y. Tretyakova. Site‐Specific 5‐Formyl Cytosine Mediated DNA‐Histone Cross‐Links: Synthesis and Polymerase Bypass by Human DNA Polymerase η. Angewandte Chemie 2021, 133 (51) , 26693-26698. https://doi.org/10.1002/ange.202109418
  25. Suresh S. Pujari, Mingxuan Wu, Jenna Thomforde, Zhipeng A. Wang, Christopher Chao, Noelle M. Olson, Luke Erber, William C. K. Pomerantz, Philip Cole, Natalia Y. Tretyakova. Site‐Specific 5‐Formyl Cytosine Mediated DNA‐Histone Cross‐Links: Synthesis and Polymerase Bypass by Human DNA Polymerase η. Angewandte Chemie International Edition 2021, 60 (51) , 26489-26494. https://doi.org/10.1002/anie.202109418
  26. Xiaoying Wei, Ying Peng, Cameron Bryan, Kun Yang. Mechanisms of DNA−protein cross-link formation and repair. Biochimica et Biophysica Acta (BBA) - Proteins and Proteomics 2021, 1869 (8) , 140669. https://doi.org/10.1016/j.bbapap.2021.140669
  27. Allison J. Matthews, Hannah M. Rowe, Jason W. Rosch, Andrew Camilli, . A Tn-seq Screen of Streptococcus pneumoniae Uncovers DNA Repair as the Major Pathway for Desiccation Tolerance and Transmission. Infection and Immunity 2021, 89 (8)https://doi.org/10.1128/IAI.00713-20
  28. Manideep C. Pachva, Alexei F. Kisselev, Bakhyt T. Matkarimov, Murat Saparbaev, Regina Groisman. DNA-Histone Cross-Links: Formation and Repair. Frontiers in Cell and Developmental Biology 2020, 8 https://doi.org/10.3389/fcell.2020.607045
  29. Leonie Hacker, Annika Dorn, Holger Puchta. WITHDRAWN: DNA-protein crosslink repair in plants. DNA Repair 2020, 88 , 102786. https://doi.org/10.1016/j.dnarep.2020.102786
  30. Leonie Hacker, Annika Dorn, Holger Puchta. Repair of DNA-protein crosslinks in plants. DNA Repair 2020, 87 , 102787. https://doi.org/10.1016/j.dnarep.2020.102787
  31. Brandon C Case, Silas Hartley, Memie Osuga, David Jeruzalmi, Manju M Hingorani. The ATPase mechanism of UvrA2 reveals the distinct roles of proximal and distal ATPase sites in nucleotide excision repair. Nucleic Acids Research 2019, 47 (8) , 4136-4152. https://doi.org/10.1093/nar/gkz180
  32. Manoj Thakur, K. Muniyappa. Nucleotide Excision Repair Pathway in Mycobacteria. 2019, 275-300. https://doi.org/10.1007/978-981-32-9413-4_16
  33. Kun Yang, Daeyoon Park, Natalia Y. Tretyakova, Marc M. Greenberg. Histone tails decrease N7-methyl-2′-deoxyguanosine depurination and yield DNA–protein cross-links in nucleosome core particles and cells. Proceedings of the National Academy of Sciences 2018, 115 (48)https://doi.org/10.1073/pnas.1813338115
  34. Lisa N. Chesner, Colin Campbell. A quantitative PCR-based assay reveals that nucleotide excision repair plays a predominant role in the removal of DNA-protein crosslinks from plasmids transfected into mammalian cells. DNA Repair 2018, 62 , 18-27. https://doi.org/10.1016/j.dnarep.2018.01.004
  35. Naeh L. Klages-Mundt, Lei Li. Formation and repair of DNA-protein crosslink damage. Science China Life Sciences 2017, 60 (10) , 1065-1076. https://doi.org/10.1007/s11427-017-9183-4
  36. Rohit Bavi, Raj Kumar, Shailima Rampogu, Minky Son, Chanin Park, Ayoung Baek, Hyong-Ha Kim, Jung-Keun Suh, Seok Ju Park, Keun Woo Lee. Molecular interactions of UvrB protein and DNA from Helicobacter pylori: Insight into a molecular modeling approach. Computers in Biology and Medicine 2016, 75 , 181-189. https://doi.org/10.1016/j.compbiomed.2016.06.005
  37. Rachel Krasich, Sunny Yang Wu, H. Kenny Kuo, Kenneth N. Kreuzer. Functions that protect Escherichia coli from DNA–protein crosslinks. DNA Repair 2015, 28 , 48-59. https://doi.org/10.1016/j.dnarep.2015.01.016
  38. A Ojha, YK Gupta. Evaluation of genotoxic potential of commonly used organophosphate pesticides in peripheral blood lymphocytes of rats. Human & Experimental Toxicology 2015, 34 (4) , 390-400. https://doi.org/10.1177/0960327114537534
  39. Susith Wickramaratne, Emily J. Boldry, Charles Buehler, Yen-Chih Wang, Mark D. Distefano, Natalia Y. Tretyakova. Error-prone Translesion Synthesis Past DNA-Peptide Cross-links Conjugated to the Major Groove of DNA via C5 of Thymidine. Journal of Biological Chemistry 2015, 290 (2) , 775-787. https://doi.org/10.1074/jbc.M114.613638
  40. Bennett Van Houten, Neil Kad. Investigation of bacterial nucleotide excision repair using single-molecule techniques. DNA Repair 2014, 20 , 41-48. https://doi.org/10.1016/j.dnarep.2013.10.012
  41. Sophie Couvé, Alexander A. Ishchenko, Olga S. Fedorova, Erlan M. Ramanculov, Jacques Laval, Murat Saparbaev, . Direct DNA Lesion Reversal and Excision Repair in Escherichia coli. EcoSal Plus 2013, 5 (2)https://doi.org/10.1128/ecosalplus.7.2.4
  42. Dessalegn B. Nemera, Amy R. Jones, Edward J. Merino. DNA Oxidation. 2013, 93-112. https://doi.org/10.1002/9781118355886.ch4
  43. Mahmoud I. Shoulkamy, Toshiaki Nakano, Makiko Ohshima, Ryoichi Hirayama, Akiko Uzawa, Yoshiya Furusawa, Hiroshi Ide. Detection of DNA–protein crosslinks (DPCs) by novel direct fluorescence labeling methods: distinct stabilities of aldehyde and radiation-induced DPCs. Nucleic Acids Research 2012, 40 (18) , e143-e143. https://doi.org/10.1093/nar/gks601
  44. Anuradha Kumari, Yun Xin Lim, Amy Hanlon Newell, Susan B. Olson, Amanda K. McCullough. Formaldehyde-induced genome instability is suppressed by an XPF-dependent pathway. DNA Repair 2012, 11 (3) , 236-246. https://doi.org/10.1016/j.dnarep.2011.11.001
  45. Kinrin Yamanaka, Irina G. Minko, Steven E. Finkel, Myron F. Goodman, R. Stephen Lloyd. Role of High-Fidelity Escherichia coli DNA Polymerase I in Replication Bypass of a Deoxyadenosine DNA-Peptide Cross-Link. Journal of Bacteriology 2011, 193 (15) , 3815-3821. https://doi.org/10.1128/JB.01550-10
  46. Hiroshi Ide, Mahmoud I. Shoulkamy, Toshiaki Nakano, Mayumi Miyamoto-Matsubara, Amir M.H. Salem. Repair and biochemical effects of DNA–protein crosslinks. Mutation Research - Fundamental and Molecular Mechanisms of Mutagenesis 2011, 711 (1-2) , 113-122. https://doi.org/10.1016/j.mrfmmm.2010.12.007
  47. Koen Wagner, Geri F. Moolenaar, Nora Goosen. Role of the insertion domain and the zinc-finger motif of Escherichia coli UvrA in damage recognition and ATP hydrolysis. DNA Repair 2011, 10 (5) , 483-496. https://doi.org/10.1016/j.dnarep.2011.02.002
  48. Jonathan T. Sczepanski, Remus S. Wong, Jeffrey N. McKnight, Gregory D. Bowman, Marc M. Greenberg. Rapid DNA–protein cross-linking and strand scission by an abasic site in a nucleosome core particle. Proceedings of the National Academy of Sciences 2010, 107 (52) , 22475-22480. https://doi.org/10.1073/pnas.1012860108
  49. Anuradha Kumari, Irina G. Minko, Rebecca L. Smith, R. Stephen Lloyd, Amanda K. McCullough. Modulation of UvrD Helicase Activity by Covalent DNA-Protein Cross-links. Journal of Biological Chemistry 2010, 285 (28) , 21313-21322. https://doi.org/10.1074/jbc.M109.078964
  50. Nora Goosen. Scanning the DNA for damage by the nucleotide excision repair machinery. DNA Repair 2010, 9 (5) , 593-596. https://doi.org/10.1016/j.dnarep.2010.02.015
  51. Aley G. Kalapila, Anthony E. Pegg. Alkyltransferase-mediated toxicity of bis-electrophiles in mammalian cells. Mutation Research - Fundamental and Molecular Mechanisms of Mutagenesis 2010, 684 (1-2) , 35-42. https://doi.org/10.1016/j.mrfmmm.2009.11.006
  52. Bendert de Graaf, Adam Clore, Amanda K. McCullough. Cellular pathways for DNA repair and damage tolerance of formaldehyde-induced DNA-protein crosslinks. DNA Repair 2009, 8 (10) , 1207-1214. https://doi.org/10.1016/j.dnarep.2009.06.007
  53. Toshiaki Nakano, Atsushi Katafuchi, Mayumi Matsubara, Hiroaki Terato, Tomohiro Tsuboi, Tasuku Masuda, Takahiro Tatsumoto, Seung Pil Pack, Keisuke Makino, Deborah L. Croteau, Bennett Van Houten, Kenta Iijima, Hiroshi Tauchi, Hiroshi Ide. Homologous Recombination but Not Nucleotide Excision Repair Plays a Pivotal Role in Tolerance of DNA-Protein Cross-links in Mammalian Cells. Journal of Biological Chemistry 2009, 284 (40) , 27065-27076. https://doi.org/10.1074/jbc.M109.019174
  54. Amir M. H. Salem, Toshiaki Nakano, Minako Takuwa, Nagisa Matoba, Tomohiro Tsuboi, Hiroaki Terato, Kazuo Yamamoto, Masami Yamada, Takehiko Nohmi, Hiroshi Ide. Genetic Analysis of Repair and Damage Tolerance Mechanisms for DNA-Protein Cross-Links in Escherichia coli. Journal of Bacteriology 2009, 191 (18) , 5657-5668. https://doi.org/10.1128/JB.00417-09
  55. L. A. Christensen, H. Wang, B. Van Houten, K. M. Vasquez. Efficient processing of TFO-directed psoralen DNA interstrand crosslinks by the UvrABC nuclease. Nucleic Acids Research 2008, 36 (22) , 7136-7145. https://doi.org/10.1093/nar/gkn880
  56. Irina G. Minko, Ivan D. Kozekov, Albena Kozekova, Thomas M. Harris, Carmelo J. Rizzo, R. Stephen Lloyd. Mutagenic potential of DNA–peptide crosslinks mediated by acrolein-derived DNA adducts. Mutation Research - Fundamental and Molecular Mechanisms of Mutagenesis 2008, 637 (1-2) , 161-172. https://doi.org/10.1016/j.mrfmmm.2007.08.001
  57. Toshiaki Nakano, Soh Morishita, Atsushi Katafuchi, Mayumi Matsubara, Yusuke Horikawa, Hiroaki Terato, Amir M.H. Salem, Shunsuke Izumi, Seung Pil Pack, Keisuke Makino, Hiroshi Ide. Nucleotide Excision Repair and Homologous Recombination Systems Commit Differentially to the Repair of DNA-Protein Crosslinks. Molecular Cell 2007, 28 (1) , 147-158. https://doi.org/10.1016/j.molcel.2007.07.029
  58. David J. Baker, Gerald Wuenschell, Liqun Xia, John Termini, Steven E. Bates, Arthur D. Riggs, Timothy R. O’Connor. Nucleotide Excision Repair Eliminates Unique DNA-Protein Cross-links from Mammalian Cells. Journal of Biological Chemistry 2007, 282 (31) , 22592-22604. https://doi.org/10.1074/jbc.M702856200
  59. Timothy R. Waters, Jitka Eryilmaz, Stella Geddes, Tracey E. Barrett. Damage detection by the UvrABC pathway: Crystal structure of UvrB bound to fluorescein‐adducted DNA. FEBS Letters 2006, 580 (27) , 6423-6427. https://doi.org/10.1016/j.febslet.2006.10.051
  60. Jung‐Suk Sung, Bruce Demple. Roles of base excision repair subpathways in correcting oxidized abasic sites in DNA. The FEBS Journal 2006, 273 (8) , 1620-1629. https://doi.org/10.1111/j.1742-4658.2006.05192.x
  61. Joyce T. Reardon, Aziz Sancar. Repair of DNA–polypeptide crosslinks by human excision nuclease. Proceedings of the National Academy of Sciences 2006, 103 (11) , 4056-4061. https://doi.org/10.1073/pnas.0600538103
  62. Bennett Van Houten, Deborah L. Croteau, Matthew J. DellaVecchia, Hong Wang, Caroline Kisker. ‘Close-fitting sleeves’: DNA damage recognition by the UvrABC nuclease system. Mutation Research - Fundamental and Molecular Mechanisms of Mutagenesis 2005, 577 (1-2) , 92-117. https://doi.org/10.1016/j.mrfmmm.2005.03.013
  63. R. Stephen Lloyd. Investigations of pyrimidine dimer glycosylases — a paradigm for DNA base excision repair enzymology. Mutation Research - Fundamental and Molecular Mechanisms of Mutagenesis 2005, 577 (1-2) , 77-91. https://doi.org/10.1016/j.mrfmmm.2005.04.003

Biochemistry

Cite this: Biochemistry 2005, 44, 8

Click to copy citationCitation copied!

Published February 1, 2005

Copyright © 2005 American Chemical Society

Altmetric

-

Citations

Learn about these metrics

Article Views are the COUNTER-compliant sum of full text article downloads since November 2008 (both PDF and HTML) across all institutions and individuals. These metrics are regularly updated to reflect usage leading up to the last few days.

Citations are the number of other articles citing this article, calculated by Crossref and updated daily. Find more information about Crossref citation counts.

The Altmetric Attention Score is a quantitative measure of the attention that a research article has received online. Clicking on the donut icon will load a page at altmetric.com with additional details about the score and the social media presence for the given article. Find more information on the Altmetric Attention Score and how the score is calculated.