{"@context":{"@vocab":"https://cir.nii.ac.jp/schema/1.0/","rdfs":"http://www.w3.org/2000/01/rdf-schema#","dc":"http://purl.org/dc/elements/1.1/","dcterms":"http://purl.org/dc/terms/","foaf":"http://xmlns.com/foaf/0.1/","prism":"http://prismstandard.org/namespaces/basic/2.0/","cinii":"http://ci.nii.ac.jp/ns/1.0/","datacite":"https://schema.datacite.org/meta/kernel-4/","ndl":"http://ndl.go.jp/dcndl/terms/","jpcoar":"https://github.com/JPCOAR/schema/blob/master/2.0/"},"@id":"https://cir.nii.ac.jp/crid/1360021390749028608.json","@type":"Article","productIdentifier":[{"identifier":{"@type":"DOI","@value":"10.1371/journal.pone.0299003"}},{"identifier":{"@type":"URI","@value":"https://dx.plos.org/10.1371/journal.pone.0299003"}},{"identifier":{"@type":"DOI","@value":"10.1101/2023.09.14.557837"}},{"identifier":{"@type":"URI","@value":"https://syndication.highwire.org/content/doi/10.1101/2023.09.14.557837"}}],"resourceType":"学術雑誌論文(journal article)","dc:title":[{"@value":"Recombinant cyclin B-Cdk1-Suc1 capable of multi-site mitotic phosphorylation in vitro"}],"description":[{"type":"abstract","notation":[{"@value":"<jats:p>Cyclin-dependent kinase 1 (Cdk1) complexed with cyclin B phosphorylates multiple sites on hundreds of proteins during mitosis. However, it is not fully understood how multi-site mitotic phosphorylation by cyclin B-Cdk1 controls the structures and functions of individual substrates. Here we develop an easy-to-use protocol to express recombinant vertebrate cyclin B and Cdk1 in insect cells from a single baculovirus vector and to purify their complexes with excellent homogeneity. A series of <jats:italic>in-vitro</jats:italic> assays demonstrate that the recombinant cyclin B-Cdk1 can efficiently and specifically phosphorylate the SP and TP motifs in substrates. The addition of Suc1 (a Cks1 homolog in fission yeast) accelerates multi-site phosphorylation of an artificial substrate containing TP motifs. Importantly, we show that mitosis-specific multi-subunit and multi-site phosphorylation of the condensin I complex can be recapitulated <jats:italic>in vitro</jats:italic> using recombinant cyclin B-Cdk1-Suc1. The materials and protocols described here will pave the way for dissecting the biochemical basis of critical mitotic processes that accompany Cdk1-mediated large-scale phosphorylation.</jats:p>"}]}],"creator":[{"@id":"https://cir.nii.ac.jp/crid/1420282801210705920","@type":"Researcher","personIdentifier":[{"@type":"KAKEN_RESEARCHERS","@value":"60462694"},{"@type":"NRID","@value":"1000060462694"},{"@type":"NRID","@value":"9000364749292"},{"@type":"NRID","@value":"9000309987878"},{"@type":"NRID","@value":"9000415140520"},{"@type":"RESEARCHMAP","@value":"https://researchmap.jp/k_shintomi"}],"foaf:name":[{"@value":"Keishi Shintomi"}]},{"@id":"https://cir.nii.ac.jp/crid/1380021390749028504","@type":"Researcher","foaf:name":[{"@value":"Yuki Masahara-Negishi"}]},{"@id":"https://cir.nii.ac.jp/crid/1380021390749028492","@type":"Researcher","foaf:name":[{"@value":"Masami Shima"}]},{"@id":"https://cir.nii.ac.jp/crid/1380021390749028608","@type":"Researcher","foaf:name":[{"@value":"Shoji Tane"}]},{"@id":"https://cir.nii.ac.jp/crid/1420564276185832448","@type":"Researcher","personIdentifier":[{"@type":"KAKEN_RESEARCHERS","@value":"50212171"},{"@type":"NRID","@value":"1000050212171"},{"@type":"NRID","@value":"9000003039059"},{"@type":"NRID","@value":"9000006065703"},{"@type":"NRID","@value":"9000018684175"},{"@type":"NRID","@value":"9000241446870"},{"@type":"NRID","@value":"9000252851458"},{"@type":"NRID","@value":"9000283917292"},{"@type":"NRID","@value":"9000008109234"},{"@type":"NRID","@value":"9000309987879"},{"@type":"NRID","@value":"9000002916904"},{"@type":"NRID","@value":"9000392060096"},{"@type":"NRID","@value":"9000017731848"},{"@type":"NRID","@value":"9000256621546"},{"@type":"NRID","@value":"9000020904146"},{"@type":"RESEARCHMAP","@value":"https://researchmap.jp/redfrog"}],"foaf:name":[{"@value":"Tatsuya Hirano"}]}],"contributor":[{"@id":"https://cir.nii.ac.jp/crid/1380021390749028624","@type":"Researcher","foaf:name":[{"@value":"Claude Prigent"}],"role":"editor"}],"publication":{"publicationIdentifier":[{"@type":"EISSN","@value":"19326203"}],"prism:publicationName":[{"@value":"PLOS ONE"}],"dc:publisher":[{"@value":"Public Library of Science (PLoS)"}],"prism:publicationDate":"2024-03-25","prism:volume":"19","prism:number":"3","prism:startingPage":"e0299003"},"reviewed":"false","dcterms:accessRights":"http://purl.org/coar/access_right/c_abf2","dc:rights":["http://creativecommons.org/licenses/by/4.0/"],"url":[{"@id":"https://dx.plos.org/10.1371/journal.pone.0299003"},{"@id":"https://syndication.highwire.org/content/doi/10.1101/2023.09.14.557837"}],"createdAt":"2024-03-25","modifiedAt":"2024-03-25","foaf:topic":[{"@id":"https://cir.nii.ac.jp/all?q=Science","dc:title":"Science"},{"@id":"https://cir.nii.ac.jp/all?q=Q","dc:title":"Q"},{"@id":"https://cir.nii.ac.jp/all?q=R","dc:title":"R"},{"@id":"https://cir.nii.ac.jp/all?q=Medicine","dc:title":"Medicine"},{"@id":"https://cir.nii.ac.jp/all?q=Research%20Article","dc:title":"Research Article"}],"project":[{"@id":"https://cir.nii.ac.jp/crid/1040018351902544000","@type":"Project","projectIdentifier":[{"@type":"KAKEN","@value":"23K23815"},{"@type":"JGN","@value":"JP23K23815"},{"@type":"URI","@value":"https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-23K23815/"}],"notation":[{"@language":"ja","@value":"分裂期染色体構築におけるクロマチン基本構造とDNAトポロジーの役割"},{"@language":"en","@value":"Roles for nucleosomes and DNA topology in mitotic chromosome assembly"}]},{"@id":"https://cir.nii.ac.jp/crid/1040282256971533952","@type":"Project","projectIdentifier":[{"@type":"KAKEN","@value":"18H05276"},{"@type":"JGN","@value":"JP18H05276"},{"@type":"URI","@value":"https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-18H05276/"}],"notation":[{"@language":"ja","@value":"コンデンシンIとIIの分子メカニズムの解明"},{"@language":"en","@value":"Molecular mechanisms of condensins I and II"}]},{"@id":"https://cir.nii.ac.jp/crid/1040285300650059136","@type":"Project","projectIdentifier":[{"@type":"KAKEN","@value":"19H05755"},{"@type":"JGN","@value":"JP19H05755"},{"@type":"URI","@value":"https://kaken.nii.ac.jp/grant/KAKENHI-PLANNED-19H05755/"}],"notation":[{"@language":"ja","@value":"核の再プログラム化のin vitro再構成"},{"@language":"en","@value":"Reconstitution of totipotent nuclei in a test tube"}]},{"@id":"https://cir.nii.ac.jp/crid/1041975275929136640","@type":"Project","projectIdentifier":[{"@type":"KAKEN","@value":"20H05938"},{"@type":"JGN","@value":"JP20H05938"},{"@type":"URI","@value":"https://kaken.nii.ac.jp/grant/KAKENHI-PLANNED-20H05938/"}],"notation":[{"@language":"ja","@value":"分裂期染色体のモダリティ"},{"@language":"en","@value":"Modality of mitotic chromosomes"}]}],"relatedProduct":[{"@id":"https://cir.nii.ac.jp/crid/1050303564713011584","@type":"Article","resourceType":"学術雑誌論文(journal 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