6RMB

Crystal structure of the DEAH-box ATPase Prp2 in complex with Spp2 and ADP

Summary for 6RMB

• Entry DOI: 10.2210/pdb6rmb/pdb
• Descriptor: Putative mRNA splicing factor, Putative pre-mRNA splicing protein, ADENOSINE-5'-DIPHOSPHATE, ... (5 entities in total)
• Functional Keywords: prp2, deah-box atpase, g-patch, spliceosome, hydrolase
• Biological source: Chaetomium thermophilum var. thermophilum DSM 1495; More
• Total number of polymer chains: 2
• Total formula weight: 76521.58

Authors

Hamann, F.,Neumann, P.,Ficner, R. (deposition date: 2019-05-06, release date: 2020-02-05, Last modification date: 2024-05-15)

Primary citation

Hamann, F.,Schmitt, A.,Favretto, F.,Hofele, R.,Neumann, P.,Xiang, S.,Urlaub, H.,Zweckstetter, M.,Ficner, R.
Structural analysis of the intrinsically disordered splicing factor Spp2 and its binding to the DEAH-box ATPase Prp2.
Proc.Natl.Acad.Sci.USA, 117:2948-2956, 2020

PubMed Abstract: The spliceosome consists of five small RNAs and more than 100 proteins. Almost 50% of the human spliceosomal proteins were predicted to be intrinsically disordered or to contain disordered regions, among them the G-patch protein Spp2. The G-patch region of Spp2 binds to the DEAH-box ATPase Prp2, and both proteins together are essential for promoting the transition from the B to the catalytically active B* spliceosome. Here we show by circular dichroism and nuclear magnetic resonance (NMR) spectroscopy that Spp2 is intrinsically disordered in solution. Crystal structures of a complex consisting of Prp2-ADP and the G-patch domain of Spp2 demonstrate that the G-patch gains a defined fold when bound to Prp2. While the N-terminal region of the G-patch always folds into an α-helix in five different crystal structures, the C-terminal part is able to adopt two alternative conformations. NMR studies further revealed that the N-terminal part of the Spp2 G-patch, which is the most conserved region in different G-patch proteins, transiently samples helical conformations, possibly facilitating a conformational selection binding mechanism. The structural analysis unveils the role of conserved residues of the G-patch in the dynamic interaction mode of Spp2 with Prp2, which is vital to maintain the binding during the Prp2 domain movements needed for RNA translocation.

PubMed: 31974312
DOI: 10.1073/pnas.1907960117

Experimental method

X-RAY DIFFRACTION (2.5 Å)