Structural and functional characterisation of human RNA helicase DHX8 provides insights into the mechanism of RNA-stimulated ADP release.
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Authors
Felisberto-Rodrigues, C
Thomas, JC
McAndrew, C
Le Bihan, Y-V
Burke, R
Workman, P
van Montfort, RLM
Thomas, JC
McAndrew, C
Le Bihan, Y-V
Burke, R
Workman, P
van Montfort, RLM
Document Type
Journal Article
Date
2019-09-13
Date Accepted
2019-08-12
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.
Citation
The Biochemical journal, 2019, 476 (18), pp. 2521 - 2543
Source Title
Publisher
PORTLAND PRESS LTD
ISSN
0264-6021
eISSN
1470-8728
Collections
Research Team
Hit Discovery & Structural Design