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dc.contributor.authorAjam, T
dc.contributor.authorDe, I
dc.contributor.authorPetkau, N
dc.contributor.authorWhelan, G
dc.contributor.authorPena, V
dc.contributor.authorEichele, G
dc.date.accessioned2020-07-03T11:19:51Z
dc.date.issued2020-06-17
dc.identifier.citationScientific reports, 2020, 10 (1), pp. 9828 - ?
dc.identifier.issn2045-2322
dc.identifier.urihttps://repository.icr.ac.uk/handle/internal/3796
dc.identifier.eissn2045-2322
dc.identifier.doi10.1038/s41598-020-66795-z
dc.description.abstractCohesin is a protein complex whose core subunits, Smc1, Smc3, Scc1, and SA1/SA2 form a ring-like structure encircling the DNA. Cohesins play a key role in the expression, repair, and segregation of eukaryotic genomes. Following a catalytic mechanism that is insufficiently understood, Esco1 and Esco2 acetyltransferases acetylate the cohesin subunit Smc3, thereby inducing stabilization of cohesin on DNA. As a prerequisite for structure-guided investigation of enzymatic activity, we determine here the crystal structure of the mouse Esco2/CoA complex at 1.8 Å resolution. We reconstitute cohesin as tri- or tetrameric assemblies and use those as physiologically-relevant substrates for enzymatic assays in vitro. Furthermore, we employ cell-based complementation studies in mouse embryonic fibroblast deficient for Esco1 and Esco2, as a means to identify catalytically-important residues in vivo. These analyses demonstrate that D567/S566 and E491/S527, located on opposite sides of the murine Esco2 active site cleft, are critical for catalysis. Our experiments support a catalytic mechanism of acetylation where residues D567 and E491 are general bases that deprotonate the ε-amino group of lysine substrate, also involving two nearby serine residues - S566 and S527- that possess a proton relay function.
dc.formatElectronic
dc.format.extent9828 - ?
dc.languageeng
dc.language.isoeng
dc.rights.urihttps://creativecommons.org/licenses/by/4.0
dc.subjectAnimals
dc.subjectHumans
dc.subjectMice
dc.subjectCoenzyme A
dc.subjectAcetyltransferases
dc.subjectChromosomal Proteins, Non-Histone
dc.subjectAmino Acid Sequence
dc.subjectCatalytic Domain
dc.subjectAcetylation
dc.subjectMutation
dc.subjectModels, Molecular
dc.subjectBiocatalysis
dc.titleAlternative catalytic residues in the active site of Esco acetyltransferases.
dc.typeJournal Article
dcterms.dateAccepted2020-05-13
rioxxterms.versionofrecord10.1038/s41598-020-66795-z
rioxxterms.licenseref.urihttps://creativecommons.org/licenses/by/4.0
rioxxterms.licenseref.startdate2020-06-17
rioxxterms.typeJournal Article/Review
dc.relation.isPartOfScientific reports
pubs.issue1
pubs.notesNot known
pubs.organisational-group/ICR
pubs.organisational-group/ICR/Primary Group
pubs.organisational-group/ICR/Primary Group/ICR Divisions
pubs.organisational-group/ICR/Primary Group/ICR Divisions/Structural Biology
pubs.organisational-group/ICR/Primary Group/ICR Divisions/Structural Biology/Mechanisms and regulation of pre-mRNA splicing
pubs.organisational-group/ICR
pubs.organisational-group/ICR/Primary Group
pubs.organisational-group/ICR/Primary Group/ICR Divisions
pubs.organisational-group/ICR/Primary Group/ICR Divisions/Structural Biology
pubs.organisational-group/ICR/Primary Group/ICR Divisions/Structural Biology/Mechanisms and regulation of pre-mRNA splicing
pubs.publication-statusPublished
pubs.volume10en_US
pubs.embargo.termsNot known
icr.researchteamMechanisms and regulation of pre-mRNA splicingen_US
dc.contributor.icrauthorPena, Vladimiren


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