dc.contributor.author | Ajam, T | |
dc.contributor.author | De, I | |
dc.contributor.author | Petkau, N | |
dc.contributor.author | Whelan, G | |
dc.contributor.author | Pena, V | |
dc.contributor.author | Eichele, G | |
dc.date.accessioned | 2020-07-03T11:19:51Z | |
dc.date.issued | 2020-06-17 | |
dc.identifier.citation | Scientific reports, 2020, 10 (1), pp. 9828 - ? | |
dc.identifier.issn | 2045-2322 | |
dc.identifier.uri | https://repository.icr.ac.uk/handle/internal/3796 | |
dc.identifier.eissn | 2045-2322 | |
dc.identifier.doi | 10.1038/s41598-020-66795-z | |
dc.description.abstract | Cohesin 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.format | Electronic | |
dc.format.extent | 9828 - ? | |
dc.language | eng | |
dc.language.iso | eng | |
dc.publisher | NATURE PUBLISHING GROUP | |
dc.rights.uri | https://creativecommons.org/licenses/by/4.0 | |
dc.subject | Animals | |
dc.subject | Humans | |
dc.subject | Mice | |
dc.subject | Coenzyme A | |
dc.subject | Acetyltransferases | |
dc.subject | Chromosomal Proteins, Non-Histone | |
dc.subject | Amino Acid Sequence | |
dc.subject | Catalytic Domain | |
dc.subject | Acetylation | |
dc.subject | Mutation | |
dc.subject | Models, Molecular | |
dc.subject | Biocatalysis | |
dc.title | Alternative catalytic residues in the active site of Esco acetyltransferases. | |
dc.type | Journal Article | |
dcterms.dateAccepted | 2020-05-13 | |
rioxxterms.versionofrecord | 10.1038/s41598-020-66795-z | |
rioxxterms.licenseref.uri | https://creativecommons.org/licenses/by/4.0 | |
rioxxterms.licenseref.startdate | 2020-06-17 | |
rioxxterms.type | Journal Article/Review | |
dc.relation.isPartOf | Scientific reports | |
pubs.issue | 1 | |
pubs.notes | Not 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-status | Published | |
pubs.volume | 10 | |
pubs.embargo.terms | Not known | |
icr.researchteam | Mechanisms and regulation of pre-mRNA splicing | |
dc.contributor.icrauthor | Pena, Vladimir | |