dc.contributor.author
Preussner, Marco
dc.contributor.author
Santos, Karine F.
dc.contributor.author
Alles, Jonathan
dc.contributor.author
Heroven, Christina
dc.contributor.author
Heyd, Florian
dc.contributor.author
Wahl, Markus C.
dc.contributor.author
Weber, Gert
dc.date.accessioned
2023-01-16T14:33:52Z
dc.date.available
2023-01-16T14:33:52Z
dc.identifier.uri
https://refubium.fu-berlin.de/handle/fub188/37628
dc.identifier.uri
http://dx.doi.org/10.17169/refubium-37343
dc.description.abstract
Small nuclear ribonucleoprotein complexes (snRNPs) represent the main subunits of the spliceosome. While the assembly of the snRNP core particles has been well characterized, comparably little is known of the incorporation of snRNP-specific proteins and the mechanisms of snRNP recycling. U5 snRNP assembly in yeast requires binding of the the Aar2 protein to Prp8p as a placeholder to preclude premature assembly of the SNRNP200 helicase, but the role of the human AAR2 homolog has not yet been investigated in detail. Here, a crystal structure of human AAR2 in complex with the RNase H-like domain of the U5-specific PRPF8 (PRP8F RH) is reported, revealing a significantly different interaction between the two proteins compared with that in yeast. Based on the structure of the AAR2–PRPF8 RH complex, the importance of the interacting regions and residues was probed and AAR2 variants were designed that failed to stably bind PRPF8 in vitro. Protein-interaction studies of AAR2 with U5 proteins using size-exclusion chromatography reveal similarities and marked differences in the interaction patterns compared with yeast Aar2p and imply phosphorylation-dependent regulation of AAR2 reminiscent of that in yeast. It is found that in vitro AAR2 seems to lock PRPF8 RH in a conformation that is only compatible with the first transesterification step of the splicing reaction and blocks a conformational switch to the step 2-like, Mg2+-coordinated conformation that is likely during U5 snRNP biogenesis. These findings extend the picture of AAR2 PRP8 interaction from yeast to humans and indicate a function for AAR2 in the spliceosomal assembly process beyond its role as an SNRNP200 placeholder in yeast.
en
dc.format.extent
11 Seiten
dc.rights.uri
https://creativecommons.org/licenses/by/4.0/
dc.subject
spliceosomal assembly
en
dc.subject.ddc
500 Naturwissenschaften und Mathematik::570 Biowissenschaften; Biologie::570 Biowissenschaften; Biologie
dc.title
Structural and functional investigation of the human snRNP assembly factor AAR2 in complex with the RNase H-like domain of PRPF8
dc.type
Wissenschaftlicher Artikel
dcterms.bibliographicCitation.doi
10.1107/S2059798322009755
dcterms.bibliographicCitation.journaltitle
Acta Crystallographica Section D: Structural Biology
dcterms.bibliographicCitation.pagestart
1373
dcterms.bibliographicCitation.pageend
1383
dcterms.bibliographicCitation.volume
78
dcterms.bibliographicCitation.url
https://doi.org/10.1107/S2059798322009755
refubium.affiliation
Biologie, Chemie, Pharmazie
refubium.affiliation.other
Institut für Chemie und Biochemie
refubium.resourceType.isindependentpub
no
dcterms.accessRights.openaire
open access
dcterms.isPartOf.eissn
2059-7983
refubium.resourceType.provider
WoS-Alert