dc.contributor.author | Felisberto-Rodrigues, C | |
dc.contributor.author | Thomas, JC | |
dc.contributor.author | McAndrew, C | |
dc.contributor.author | Le Bihan, Y-V | |
dc.contributor.author | Burke, R | |
dc.contributor.author | Workman, P | |
dc.contributor.author | van Montfort, RLM | |
dc.date.accessioned | 2020-05-28T12:56:07Z | |
dc.date.issued | 2019-09-13 | |
dc.identifier.citation | The Biochemical journal, 2019, 476 (18), pp. 2521 - 2543 | |
dc.identifier.issn | 0264-6021 | |
dc.identifier.uri | https://repository.icr.ac.uk/handle/internal/3661 | |
dc.identifier.eissn | 1470-8728 | |
dc.identifier.doi | 10.1042/bcj20190383 | |
dc.description.abstract | DHX8 is a crucial DEAH-box RNA helicase involved in splicing and required for the release of mature mRNA from the spliceosome. Here, we report the biochemical characterisation of full-length human DHX8 and the catalytically active helicase core DHX8Δ547, alongside crystal structures of DHX8Δ547 bound to ADP and a structure of DHX8Δ547 bound to poly(A)6 single-strand RNA. Our results reveal that DHX8 has an in vitro binding preference for adenine-rich RNA and that RNA binding triggers the release of ADP through significant conformational flexibility in the conserved DEAH-, P-loop and hook-turn motifs. We demonstrate the importance of R620 and both the hook-turn and hook-loop regions for DHX8 helicase activity and propose that the hook-turn acts as a gatekeeper to regulate the directional movement of the 3' end of RNA through the RNA-binding channel. This study provides an in-depth understanding of the activity of DHX8 and contributes insights into the RNA-unwinding mechanisms of the DEAH-box helicase family. | |
dc.format | Electronic | |
dc.format.extent | 2521 - 2543 | |
dc.language | eng | |
dc.language.iso | eng | |
dc.publisher | PORTLAND PRESS LTD | |
dc.rights.uri | https://creativecommons.org/licenses/by/4.0 | |
dc.subject | Humans | |
dc.subject | RNA | |
dc.subject | Poly A | |
dc.subject | Adenosine Diphosphate | |
dc.subject | Amino Acid Motifs | |
dc.subject | Protein Binding | |
dc.subject | Structure-Activity Relationship | |
dc.subject | DEAD-box RNA Helicases | |
dc.subject | RNA Splicing Factors | |
dc.title | Structural and functional characterisation of human RNA helicase DHX8 provides insights into the mechanism of RNA-stimulated ADP release. | |
dc.type | Journal Article | |
dcterms.dateAccepted | 2019-08-12 | |
rioxxterms.versionofrecord | 10.1042/bcj20190383 | |
rioxxterms.licenseref.uri | https://creativecommons.org/licenses/by/4.0 | |
rioxxterms.licenseref.startdate | 2019-09-13 | |
rioxxterms.type | Journal Article/Review | |
dc.relation.isPartOf | The Biochemical journal | |
pubs.issue | 18 | |
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/Cancer Therapeutics | |
pubs.organisational-group | /ICR/Primary Group/ICR Divisions/Cancer Therapeutics/Hit Discovery & Structural Design | |
pubs.organisational-group | /ICR/Primary Group/ICR Divisions/Structural Biology | |
pubs.organisational-group | /ICR/Primary Group/ICR Divisions/Structural Biology/Hit Discovery & Structural Design | |
pubs.publication-status | Published | |
pubs.volume | 476 | |
pubs.embargo.terms | Not known | |
icr.researchteam | Hit Discovery & Structural Design | |
dc.contributor.icrauthor | Le Bihan, Yann-Vai | |
dc.contributor.icrauthor | Burke, Rosemary | |
dc.contributor.icrauthor | Workman, Paul | |
dc.contributor.icrauthor | Van Montfort, Robert | |