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  Prp2-mediated protein rearrangements at the catalytic core of the spliceosome as revealed by dcFCCS.

Ohrt, T., Prior, M., Dannenberg, J., Odenwälder, P., Dybkov, O., Rasche, N., et al. (2012). Prp2-mediated protein rearrangements at the catalytic core of the spliceosome as revealed by dcFCCS. RNA, 18(6), 1244-1256. doi:10.1261/rna.033316.112.

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Item Permalink: http://hdl.handle.net/11858/00-001M-0000-000F-A623-6 Version Permalink: http://hdl.handle.net/11858/00-001M-0000-0027-C6A1-E
Genre: Journal Article

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 Creators:
Ohrt, T.1, Author              
Prior, M., Author
Dannenberg, J.1, Author              
Odenwälder, P.1, Author              
Dybkov, O.1, Author              
Rasche, N.1, Author              
Schmitzova, J.1, Author              
Gregor, I., Author
Fabrizio, P.1, Author              
Enderlein, J., Author
Lührmann, R.1, Author              
Affiliations:
1Department of Cellular Biochemistry, MPI for biophysical chemistry, Max Planck Society, ou_578576              

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Free keywords: Cwc24; dcFCCS; Prp2-mediated; spliceosome dynamics; U2 SF3a/b
 Abstract: The compositional and conformational changes during catalytic activation of the spliceosome promoted by the DEAH box ATPase Prp2 are only poorly understood. Here, we show by dual-color fluorescence cross-correlation spectroscopy (dcFCCS) that the binding affinity of several proteins is significantly changed during the Prp2-mediated transition of precatalytic Bact spliceosomes to catalytically activated B* spliceosomes from Saccharomyces cerevisiae. During this step, several proteins, including the zinc-finger protein Cwc24, are quantitatively displaced from the B* complex. Consistent with this, we show that Cwc24 is required for step 1 but not for catalysis per se. The U2-associated SF3a and SF3b proteins Prp11 and Cus1 remain bound to the B* spliceosome under near-physiological conditions, but their binding is reduced at high salt. Conversely, high-affinity binding sites are created for Yju2 and Cwc25 during catalytic activation, consistent with their requirement for step 1 catalysis. Our results suggest high cooperativity of multiple Prp2-mediated structural rearrangements at the spliceosome’s catalytic core. Moreover, dcFCCS represents a powerful tool ideally suited to study quantitatively spliceosomal protein dynamics in equilibrium.

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Language(s): eng - English
 Dates: 2012-04-252012
 Publication Status: Published in print
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 Rev. Method: Peer
 Identifiers: DOI: 10.1261/rna.033316.112
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Title: RNA
Source Genre: Journal
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Pages: - Volume / Issue: 18 (6) Sequence Number: - Start / End Page: 1244 - 1256 Identifier: -