| Autor: |
Felisberto-Rodrigues, C., Thomas, J.C., McAndrew, C., Le Bihan, Y.V., Burke, R., Workman, P., van Montfort, R.L.M. |
| Jazyk: |
angličtina |
| Rok vydání: |
2019 |
| Zdroj: |
'Biochemical Journal ', vol: 476, pages: 2521-2543 (2019) |
| ISSN: |
0264-6021 |
| Popis: |
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 DHX8 Delta 547, alongside crystal structures of DHX8 Delta 547 bound to ADP and a structure of DHX8 Delta 547 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. |
| Databáze: |
OpenAIRE |
| Externí odkaz: |
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