{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,3,24]],"date-time":"2026-03-24T22:04:02Z","timestamp":1774389842679,"version":"3.50.1"},"reference-count":57,"publisher":"MDPI AG","issue":"1","license":[{"start":{"date-parts":[[2022,12,21]],"date-time":"2022-12-21T00:00:00Z","timestamp":1671580800000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"funder":[{"name":"FCT","award":["POCTI\/32621\/BME\/2000"],"award-info":[{"award-number":["POCTI\/32621\/BME\/2000"]}]},{"name":"FCT","award":["POCI\/BIA-PRO\/58344\/2004"],"award-info":[{"award-number":["POCI\/BIA-PRO\/58344\/2004"]}]},{"name":"FCT","award":["PTDC\/QUI-BIQ\/098427\/2008"],"award-info":[{"award-number":["PTDC\/QUI-BIQ\/098427\/2008"]}]},{"name":"FCT","award":["PEst-OE\/EQB\/LA0023\/2011"],"award-info":[{"award-number":["PEst-OE\/EQB\/LA0023\/2011"]}]},{"name":"FCT","award":["Do 649\/6-1"],"award-info":[{"award-number":["Do 649\/6-1"]}]},{"name":"Deutsche Forschungsgemeinschaft","award":["POCTI\/32621\/BME\/2000"],"award-info":[{"award-number":["POCTI\/32621\/BME\/2000"]}]},{"name":"Deutsche Forschungsgemeinschaft","award":["POCI\/BIA-PRO\/58344\/2004"],"award-info":[{"award-number":["POCI\/BIA-PRO\/58344\/2004"]}]},{"name":"Deutsche Forschungsgemeinschaft","award":["PTDC\/QUI-BIQ\/098427\/2008"],"award-info":[{"award-number":["PTDC\/QUI-BIQ\/098427\/2008"]}]},{"name":"Deutsche Forschungsgemeinschaft","award":["PEst-OE\/EQB\/LA0023\/2011"],"award-info":[{"award-number":["PEst-OE\/EQB\/LA0023\/2011"]}]},{"name":"Deutsche Forschungsgemeinschaft","award":["Do 649\/6-1"],"award-info":[{"award-number":["Do 649\/6-1"]}]}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":["Biomolecules"],"abstract":"<jats:p>Whereas assembly of the 20S proteasome core particle (CP) in prokaryotes apparently occurs spontaneously, the efficiency of this process in eukaryotes relies on the dedicated assembly chaperones Ump1, Pba1-Pba2, and Pba3-Pba4. For mammals, it was reported that CP assembly initiates with formation of a complete \u03b1-ring that functions as a template for \u03b2 subunit incorporation. By contrast, we were not able to detect a ring composed only of a complete set of \u03b1 subunits in S. cerevisiae. Instead, we found that the CP subunits \u03b11, \u03b12, and \u03b14 each form independent small complexes. Purification of such complexes containing \u03b14 revealed the presence of chaperones of the Hsp70\/Ssa and Hsp110\/Sse families. Consistently, certain small complexes containing \u03b11, \u03b12, and \u03b14 were not formed in strains lacking these chaperones. Deletion of the SSE1 gene in combination with deletions of PRE9 (\u03b13), PBA3, or UMP1 genes resulted in severe synthetic growth defects, high levels of ubiquitin-conjugates, and an accumulation of distinct small complexes with \u03b1 subunits. Our study shows that Hsp70 and Hsp110 chaperones cooperate to promote the folding of individual \u03b1 subunits and\/or their assembly with other CP subunits, Ump1, and Pba1-Pba4 in subsequent steps.<\/jats:p>","DOI":"10.3390\/biom13010011","type":"journal-article","created":{"date-parts":[[2022,12,21]],"date-time":"2022-12-21T05:42:53Z","timestamp":1671601373000},"page":"11","update-policy":"https:\/\/doi.org\/10.3390\/mdpi_crossmark_policy","source":"Crossref","is-referenced-by-count":8,"title":["Hsp70 and Hsp110 Chaperones Promote Early Steps of Proteasome Assembly"],"prefix":"10.3390","volume":"13","author":[{"ORCID":"https:\/\/orcid.org\/0000-0002-6385-9117","authenticated-orcid":false,"given":"Ana C.","family":"Matias","sequence":"first","affiliation":[{"name":"Center of Molecular Biosciences, Institute for Genetics, Department of Biology, Faculty of Natural Sciences and Mathematics, University of Cologne, 50674 Cologne, Germany"},{"name":"Departamento de Qu\u00edmica e Bioqu\u00edmica, Faculdade de Ci\u00eancias e Tecnologia, Universidade do Algarve, 8000-117 Faro, Portugal"}]},{"given":"Joao","family":"Matos","sequence":"additional","affiliation":[{"name":"Departamento de Qu\u00edmica e Bioqu\u00edmica, Faculdade de Ci\u00eancias e Tecnologia, Universidade do Algarve, 8000-117 Faro, Portugal"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-5756-6780","authenticated-orcid":false,"given":"R. J\u00fcrgen","family":"Dohmen","sequence":"additional","affiliation":[{"name":"Center of Molecular Biosciences, Institute for Genetics, Department of Biology, Faculty of Natural Sciences and Mathematics, University of Cologne, 50674 Cologne, Germany"}]},{"given":"Paula C.","family":"Ramos","sequence":"additional","affiliation":[{"name":"Center of Molecular Biosciences, Institute for Genetics, Department of Biology, Faculty of Natural Sciences and Mathematics, University of Cologne, 50674 Cologne, Germany"},{"name":"Departamento de Qu\u00edmica e Bioqu\u00edmica, Faculdade de Ci\u00eancias e Tecnologia, Universidade do Algarve, 8000-117 Faro, Portugal"}]}],"member":"1968","published-online":{"date-parts":[[2022,12,21]]},"reference":[{"key":"ref_1","doi-asserted-by":"crossref","first-page":"1242","DOI":"10.1038\/nm.3739","article-title":"Ubiquitination in disease pathogenesis and treatment","volume":"20","author":"Popovic","year":"2014","journal-title":"Nat. Med."},{"key":"ref_2","doi-asserted-by":"crossref","unstructured":"Goetzke, C.C., Ebstein, F., and Kallinich, T. (2021). Role of proteasomes in inflammation. J. Clin. Med., 10.","DOI":"10.3390\/jcm10081783"},{"key":"ref_3","doi-asserted-by":"crossref","first-page":"319","DOI":"10.1534\/genetics.112.140467","article-title":"The ubiquitin-proteasome system of Saccharomyces cerevisiae","volume":"192","author":"Finley","year":"2012","journal-title":"Genetics"},{"key":"ref_4","doi-asserted-by":"crossref","first-page":"13","DOI":"10.1016\/j.bbamcr.2013.08.012","article-title":"Regulation of proteasome activity in health and disease","volume":"1843","author":"Schmidt","year":"2014","journal-title":"Biochim. Biophys. Acta"},{"key":"ref_5","doi-asserted-by":"crossref","first-page":"165793","DOI":"10.1016\/j.bbadis.2020.165793","article-title":"Defective proteasome biogenesis into skin fibroblasts isolated from Rett syndrome subjects with MeCP2 non-sense mutations","volume":"1866","author":"Sbardella","year":"2020","journal-title":"Biochim. Biophys. Acta Mol. Basis Dis."},{"key":"ref_6","doi-asserted-by":"crossref","first-page":"3458","DOI":"10.1158\/0008-5472.CAN-17-2296","article-title":"PSMD5 Inactivation promotes 26S proteasome assembly during colorectal tumor progression","volume":"78","author":"Levin","year":"2018","journal-title":"Cancer Res."},{"key":"ref_7","doi-asserted-by":"crossref","first-page":"603","DOI":"10.1016\/j.celrep.2012.07.013","article-title":"Role of S5b\/PSMD5 in proteasome inhibition caused by TNF-\u03b1\/NF\u03baB in higher eukaryotes","volume":"2","author":"Shim","year":"2012","journal-title":"Cell Rep."},{"key":"ref_8","doi-asserted-by":"crossref","first-page":"887","DOI":"10.1016\/j.cell.2009.04.061","article-title":"Multiple assembly chaperones govern biogenesis of the proteasome regulatory particle base","volume":"137","author":"Funakoshi","year":"2009","journal-title":"Cell"},{"key":"ref_9","doi-asserted-by":"crossref","first-page":"1296","DOI":"10.1016\/j.str.2008.07.001","article-title":"PACemakers of proteasome core particle assembly","volume":"16","author":"Ramos","year":"2008","journal-title":"Structure"},{"key":"ref_10","doi-asserted-by":"crossref","first-page":"104","DOI":"10.1038\/nrm2630","article-title":"Molecular mechanisms of proteasome assembly","volume":"10","author":"Murata","year":"2009","journal-title":"Nat. Rev. Mol. Cell Biol."},{"key":"ref_11","doi-asserted-by":"crossref","first-page":"415","DOI":"10.1146\/annurev-biochem-060410-150257","article-title":"Molecular architecture and assembly of the eukaryotic proteasome","volume":"82","author":"Tomko","year":"2013","journal-title":"Annu. Rev. Biochem."},{"key":"ref_12","doi-asserted-by":"crossref","first-page":"463","DOI":"10.1038\/386463a0","article-title":"Structure of 20S proteasome from yeast at 2.4 A resolution","volume":"386","author":"Groll","year":"1997","journal-title":"Nature"},{"key":"ref_13","doi-asserted-by":"crossref","first-page":"500","DOI":"10.1038\/sj.emboj.7600059","article-title":"Plasticity in eucaryotic 20S proteasome ring assembly revealed by a subunit deletion in yeast","volume":"23","author":"Velichutina","year":"2004","journal-title":"EMBO J."},{"key":"ref_14","doi-asserted-by":"crossref","first-page":"2962","DOI":"10.1016\/j.celrep.2016.02.068","article-title":"Assembly of an evolutionarily conserved alternative proteasome isoform in human cells","volume":"14","author":"Padmanabhan","year":"2016","journal-title":"Cell Rep."},{"key":"ref_15","doi-asserted-by":"crossref","first-page":"17","DOI":"10.1016\/j.tibs.2013.10.004","article-title":"The unique functions of tissue-specific proteasomes","volume":"39","author":"Kniepert","year":"2014","journal-title":"Trends Biochem. Sci."},{"key":"ref_16","doi-asserted-by":"crossref","first-page":"489","DOI":"10.1016\/S0092-8674(00)80942-3","article-title":"Ump1p is required for proper maturation of the 20S proteasome and becomes its substrate upon completion of the assembly","volume":"92","author":"Ramos","year":"1998","journal-title":"Cell"},{"key":"ref_17","doi-asserted-by":"crossref","first-page":"2339","DOI":"10.1038\/sj.emboj.7601681","article-title":"beta-Subunit appendages promote 20S proteasome assembly by overcoming an Ump1-dependent checkpoint","volume":"26","author":"Li","year":"2007","journal-title":"EMBO J."},{"key":"ref_18","doi-asserted-by":"crossref","first-page":"660","DOI":"10.1016\/j.molcel.2007.06.025","article-title":"20S proteasome assembly is orchestrated by two distinct pairs of chaperones in yeast and in mammals","volume":"27","author":"Barrault","year":"2007","journal-title":"Mol. Cell"},{"key":"ref_19","doi-asserted-by":"crossref","first-page":"6123","DOI":"10.1038\/ncomms7123","article-title":"Proteasome assembly from 15S precursors involves major conformational changes and recycling of the Pba1-Pba2 chaperone","volume":"6","author":"Kock","year":"2015","journal-title":"Nat. Commun."},{"key":"ref_20","doi-asserted-by":"crossref","first-page":"418","DOI":"10.1038\/s41594-021-00583-9","article-title":"Structures of chaperone-associated assembly intermediates reveal coordinated mechanisms of proteasome biogenesis","volume":"28","author":"Schnell","year":"2021","journal-title":"Nat. Struct. Mol. Biol."},{"key":"ref_21","doi-asserted-by":"crossref","first-page":"34869","DOI":"10.1074\/jbc.M705836200","article-title":"The C-terminal extension of the beta7 subunit and activator complexes stabilize nascent 20 S proteasomes and promote their maturation","volume":"282","author":"Marques","year":"2007","journal-title":"J. Biol. Chem."},{"key":"ref_22","doi-asserted-by":"crossref","unstructured":"Zimmermann, J., Ramos, P.C., and Dohmen, R.J. (2022). Interaction with the assembly chaperone Ump1 promotes incorporation of the \u03b27 subunit into half-proteasome precursor complexes driving their dimerization. Biomolecules, 12.","DOI":"10.3390\/biom12020253"},{"key":"ref_23","doi-asserted-by":"crossref","first-page":"765","DOI":"10.1038\/nsb1194-765","article-title":"Critical elements in proteasome assembly","volume":"1","author":"Zwickl","year":"1994","journal-title":"Nat. Struct. Biol."},{"key":"ref_24","first-page":"418189","article-title":"Dissecting the assembly pathway of the 20S proteasome","volume":"24","author":"Golbik","year":"1997","journal-title":"FEBS Lett."},{"key":"ref_25","doi-asserted-by":"crossref","first-page":"13130","DOI":"10.1038\/srep13130","article-title":"Alpha-ring independent assembly of the 20S proteasome","volume":"19","author":"Panfair","year":"2015","journal-title":"Sci. Rep."},{"key":"ref_26","doi-asserted-by":"crossref","first-page":"1381","DOI":"10.1038\/nature04106","article-title":"A heterodimeric complex that promotes the assembly of mammalian 20S proteasomes","volume":"437","author":"Hirano","year":"2005","journal-title":"Nature"},{"key":"ref_27","doi-asserted-by":"crossref","first-page":"977","DOI":"10.1016\/j.molcel.2006.11.015","article-title":"Cooperation of multiple chaperones required for the assembly of mammalian 20S proteasomes","volume":"24","author":"Hirano","year":"2006","journal-title":"Mol. Cell"},{"key":"ref_28","doi-asserted-by":"crossref","first-page":"107","DOI":"10.1016\/j.bbrc.2020.02.044","article-title":"A novel proteasome assembly intermediate bypasses the need to form \u03b1-rings first","volume":"525","author":"Hammack","year":"2020","journal-title":"Biochem. Biophys. Res. Commun."},{"key":"ref_29","doi-asserted-by":"crossref","first-page":"18099","DOI":"10.1074\/jbc.270.30.18099","article-title":"An essential yeast gene encoding a homolog of ubiquitin-activating enzyme","volume":"270","author":"Dohmen","year":"1995","journal-title":"J. Biol. Chem."},{"key":"ref_30","doi-asserted-by":"crossref","first-page":"953","DOI":"10.1002\/(SICI)1097-0061(199807)14:10<953::AID-YEA293>3.0.CO;2-U","article-title":"Additional modules for versatile and economical PCR-based gene deletion and modification in Saccharomyces cerevisiae","volume":"14","author":"Longtine","year":"1998","journal-title":"Yeast"},{"key":"ref_31","doi-asserted-by":"crossref","first-page":"527","DOI":"10.1016\/0378-1119(88)90185-0","article-title":"New yeast-Escherichia coli shuttle vectors constructed with in vitro mutagenized yeast genes lacking six- base pair restriction sites","volume":"74","author":"Gietz","year":"1988","journal-title":"Gene"},{"key":"ref_32","doi-asserted-by":"crossref","first-page":"5767","DOI":"10.1093\/nar\/22.25.5767","article-title":"Regulatable promoters of Saccharomyces cerevisiae: Comparison of transcriptional activity and their use for heterologous expression","volume":"22","author":"Mumberg","year":"1994","journal-title":"Nucleic Acids Res."},{"key":"ref_33","doi-asserted-by":"crossref","first-page":"947","DOI":"10.1002\/yea.1142","article-title":"A versatile toolbox for PCR-based tagging of yeast genes: New fluorescent proteins, more markers and promoter substitution cassettes","volume":"21","author":"Janke","year":"2004","journal-title":"Yeast"},{"key":"ref_34","doi-asserted-by":"crossref","first-page":"963","DOI":"10.1002\/(SICI)1097-0061(199907)15:10B<963::AID-YEA399>3.0.CO;2-W","article-title":"Epitope tagging of yeast genes using a PCR-based strategy: More tags and improved practical routines","volume":"15","author":"Knop","year":"1999","journal-title":"Yeast"},{"key":"ref_35","doi-asserted-by":"crossref","first-page":"228","DOI":"10.1038\/nsmb.1386","article-title":"Crystal structure of a chaperone complex that contributes to the assembly of yeast 20S proteasomes","volume":"15","author":"Yashiroda","year":"2008","journal-title":"Nat. Struct. Mol. Biol."},{"key":"ref_36","doi-asserted-by":"crossref","first-page":"1","DOI":"10.1379\/CSC-245R.1","article-title":"All in the family: Atypical Hsp70 chaperones are conserved modulators of Hsp70 activity","volume":"12","author":"Shaner","year":"2007","journal-title":"Cell Stress Chaperones."},{"key":"ref_37","doi-asserted-by":"crossref","first-page":"321","DOI":"10.1016\/j.molcel.2010.07.012","article-title":"Gymnastics of molecular chaperones","volume":"39","author":"Mayer","year":"2010","journal-title":"Mol. Cell"},{"key":"ref_38","doi-asserted-by":"crossref","first-page":"665","DOI":"10.1038\/s41580-019-0133-3","article-title":"The Hsp70 chaperone network","volume":"20","author":"Rosenzweig","year":"2019","journal-title":"Nat. Rev. Mol. Cell Biol."},{"key":"ref_39","doi-asserted-by":"crossref","first-page":"57","DOI":"10.1016\/0378-1119(93)90514-4","article-title":"Isolation and characterization of SSE1 and SSE2, new members of the yeast HSP70 multigene family","volume":"132","author":"Mukai","year":"1993","journal-title":"Gene"},{"key":"ref_40","doi-asserted-by":"crossref","first-page":"41262","DOI":"10.1074\/jbc.M503614200","article-title":"The yeast Hsp110 Sse1 functionally interacts with the Hsp70 chaperones Ssa and Ssb","volume":"280","author":"Shaner","year":"2005","journal-title":"J. Biol. Chem."},{"key":"ref_41","doi-asserted-by":"crossref","first-page":"2519","DOI":"10.1038\/sj.emboj.7601138","article-title":"Molecular chaperones of the Hsp110 family act as nucleotide exchange factors of Hsp70s","volume":"25","author":"Dragovic","year":"2006","journal-title":"EMBO J."},{"key":"ref_42","doi-asserted-by":"crossref","first-page":"2510","DOI":"10.1038\/sj.emboj.7601139","article-title":"Chaperone network in the yeast cytosol: Hsp110 is revealed as an Hsp70 nucleotide exchange factor","volume":"25","author":"Raviol","year":"2006","journal-title":"EMBO J."},{"key":"ref_43","doi-asserted-by":"crossref","first-page":"31636","DOI":"10.1074\/jbc.272.50.31636","article-title":"Hsp110 protects heat-denatured proteins and confers cellular thermoresistance","volume":"272","author":"Oh","year":"1997","journal-title":"J. Biol. Chem."},{"key":"ref_44","doi-asserted-by":"crossref","first-page":"15712","DOI":"10.1074\/jbc.274.22.15712","article-title":"The chaperoning activity of Hsp110. Identification of functional domains by use of targeted deletions","volume":"274","author":"Oh","year":"1999","journal-title":"J. Biol. Chem."},{"key":"ref_45","doi-asserted-by":"crossref","first-page":"95","DOI":"10.1006\/jmbi.1997.0947","article-title":"Maturation of mammalian 20 S proteasome: Purification and characterization of 13 S and 16 S proteasome precursor complexes","volume":"268","author":"Schmidtke","year":"1997","journal-title":"J. Mol. Biol."},{"key":"ref_46","doi-asserted-by":"crossref","first-page":"438","DOI":"10.1016\/j.bbrc.2017.03.059","article-title":"Molecular chaperones of the Hsp70 family assist in the assembly of 20S proteasomes","volume":"486","author":"Hammack","year":"2017","journal-title":"Biochem. Biophys. Res. Commun."},{"key":"ref_47","doi-asserted-by":"crossref","first-page":"164","DOI":"10.1016\/j.bbrc.2016.11.024","article-title":"Assembly of proteasome subunits into non-canonical complexes in vivo","volume":"482","author":"Hammack","year":"2017","journal-title":"Biochem. Biophys. Res. Commun."},{"key":"ref_48","doi-asserted-by":"crossref","first-page":"21992","DOI":"10.1074\/jbc.M313739200","article-title":"The function of the yeast molecular chaperone Sse1 is mechanistically distinct from the closely related hsp70 family","volume":"279","author":"Shaner","year":"2004","journal-title":"J. Biol. Chem."},{"key":"ref_49","first-page":"2568","article-title":"Complex interactions among members of an essential subfamily of hsp70 genes in Saccharomyces cerevisiae","volume":"7","author":"Stone","year":"1987","journal-title":"Mol. Cell Biol."},{"key":"ref_50","doi-asserted-by":"crossref","first-page":"15075","DOI":"10.1021\/bi061279k","article-title":"Characterization of Hsp70 binding and nucleotide exchange by the yeast Hsp110 chaperone Sse1","volume":"45","author":"Shaner","year":"2006","journal-title":"Biochemistry"},{"key":"ref_51","doi-asserted-by":"crossref","first-page":"232","DOI":"10.1016\/j.molcel.2008.05.006","article-title":"Structure of the Hsp110:Hsc70 nucleotide exchange machine","volume":"31","author":"Schuermann","year":"2008","journal-title":"Mol. Cell"},{"key":"ref_52","doi-asserted-by":"crossref","first-page":"774","DOI":"10.1016\/j.jmb.2007.05.022","article-title":"Inference of macromolecular assemblies from crystalline state","volume":"372","author":"Krissinel","year":"2007","journal-title":"J. Mol. Biol."},{"key":"ref_53","doi-asserted-by":"crossref","first-page":"1067","DOI":"10.1016\/j.febslet.2013.02.018","article-title":"The control of spindle length by Hsp70 and Hsp110 molecular chaperones","volume":"587","author":"Makhnevych","year":"2013","journal-title":"FEBS Lett."},{"key":"ref_54","doi-asserted-by":"crossref","first-page":"1110","DOI":"10.1016\/j.bbrc.2014.06.119","article-title":"Pba3-Pba4 heterodimer acts as a molecular matchmaker in proteasome alpha-ring formation","volume":"450","author":"Takagi","year":"2014","journal-title":"Biochem. Biophys. Res. Commun."},{"key":"ref_55","doi-asserted-by":"crossref","first-page":"349","DOI":"10.1042\/bj3440349","article-title":"alpha5 subunit in Trypanosoma brucei proteasome can self-assemble to form a cylinder of four stacked heptamer rings","volume":"344","author":"Yao","year":"1999","journal-title":"Biochem. J."},{"key":"ref_56","doi-asserted-by":"crossref","first-page":"10080","DOI":"10.1074\/jbc.272.15.10080","article-title":"The human alpha-type proteasomal subunit HsC8 forms a double ring like structure, but does not assemble into proteasome-like particles with the beta-type subunits HsDelta or HsBPROS26","volume":"272","author":"Gerards","year":"1997","journal-title":"J. Biol. Chem."},{"key":"ref_57","doi-asserted-by":"crossref","first-page":"21399","DOI":"10.1074\/jbc.M113.479253","article-title":"Hsp110 is a Bona Fide chaperone using ATP to unfold stable misfolded polypeptides and reciprocally collaborate with Hsp70 to solubilize protein aggregates","volume":"288","author":"Mattoo","year":"2013","journal-title":"J. Biol. Chem."}],"container-title":["Biomolecules"],"original-title":[],"language":"en","link":[{"URL":"https:\/\/www.mdpi.com\/2218-273X\/13\/1\/11\/pdf","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2025,10,11]],"date-time":"2025-10-11T01:45:17Z","timestamp":1760147117000},"score":1,"resource":{"primary":{"URL":"https:\/\/www.mdpi.com\/2218-273X\/13\/1\/11"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2022,12,21]]},"references-count":57,"journal-issue":{"issue":"1","published-online":{"date-parts":[[2023,1]]}},"alternative-id":["biom13010011"],"URL":"https:\/\/doi.org\/10.3390\/biom13010011","relation":{},"ISSN":["2218-273X"],"issn-type":[{"value":"2218-273X","type":"electronic"}],"subject":[],"published":{"date-parts":[[2022,12,21]]}}}