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. 2019 Dec 10;48(3):1508–1530. doi: 10.1093/nar/gkz1148

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© The Author(s) 2019. Published by Oxford University Press on behalf of Nucleic Acids Research.

This is an Open Access article distributed under the terms of the Creative Commons Attribution License (http://creativecommons.org/licenses/by/4.0/), which permits unrestricted reuse, distribution, and reproduction in any medium, provided the original work is properly cited.

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Figure 7. — In vitro assembly of the U7-specific ring on a spliceosomal Sm site. (A) Nucleotide sequence of the U7-specific Sm site (top sequence) and changes made to generate C-G and spliceosomal-type (Spl) Sm sites (bottom sequences). Mutated nucleotides are indicated with the arrows. (B) Interaction between the spliceosomal Sm proteins and individual nucleotides of the Sm site in U4 snRNA (50). (C) Proteins of the U7-specific Sm ring bound to the U7, C-G and Spl Sm sites visualized by silver staining (top) and Western blotting (bottom). (D) Processing of ΔSL/Ys pre-mRNA in a mouse nuclear extract in the presence of 1, 3 and 5 μl of peak fraction containing indicated core U7 snRNPs. The arrow denotes the unusual cleavage product generated by SupU7 snRNP containing Spl Sm site. (E, F) Processing of ΔSL/Ys (panel E) and H2a WT/Ys (panel F) pre-mRNAs in a mouse nuclear extract in the presence of 5 or 2 μl of indicated core SupU7 snRNPs and processing competitors (SL or αSupU7).