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Journal Article

Reconstitution of the human U snRNP assembly machinery reveals stepwise Sm protein organization.

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Chari,  A.
Research Group of Structural Biochemistry and Mechanisms, MPI for Biophysical Chemistry, Max Planck Society;

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2166779_Suppl_1.pdf
(Supplementary material), 8MB

2166779_Suppl_2.pdf
(Supplementary material), 26MB

2166779_Suppl_3.pdf.part
(Supplementary material), 25MB

2166779_Suppl_4.pdf
(Supplementary material), 549KB

2166779_Suppl_5.pdf
(Supplementary material), 9MB

2166779_Suppl_6.pdf
(Supplementary material), 5MB

2166779_Suppl_7.pdf
(Supplementary material), 197KB

2166779_Suppl_8.pdf
(Supplementary material), 257KB

2166779_Suppl_9.pdf
(Supplementary material), 75KB

2166779_Suppl_10.docx
(Supplementary material), 28KB

Citation

Neuenkirchen, N., Englbrecht, C., Ohmer, J., Ziegenhals, T., Chari, A., & Fischer, U. (2015). Reconstitution of the human U snRNP assembly machinery reveals stepwise Sm protein organization. EMBO Journal, 34(14), 1925-1941. doi:10.15252/embj.201490350.


Cite as: https://hdl.handle.net/11858/00-001M-0000-0027-A83A-A
Abstract
The assembly of spliceosomal U snRNPs depends on the coordinated action of PRMT5 and SMN complexes in vivo. These trans-acting factors enable the faithful delivery of seven Sm proteins onto snRNA and the formation of the common core of snRNPs. To gain mechanistic insight into their mode of action, we reconstituted the assembly machinery from recombinant sources. We uncover a stepwise and ordered formation of distinct Sm protein complexes on the PRMT5 complex, which is facilitated by the assembly chaperone pICln. Upon completion, the formed pICln-Sm units are displaced by new pICln-Sm protein substrates and transferred onto the SMN complex. The latter acts as a Brownian machine that couples spontaneous conformational changes driven by thermal energy to prevent mis-assembly and to ensure the transfer of Sm proteins to cognate RNA. Investigation of mutant SMN complexes provided insight into the contribution of individual proteins to these activities. The biochemical reconstitution presented here provides a basis for a detailed molecular dissection of the U snRNP assembly reaction.