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. 2019 Nov;25(11):1509–1521. doi: 10.1261/rna.072256.119

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© 2019 Martelly et al.; Published by Cold Spring Harbor Laboratory Press for the RNA Society

This article is distributed exclusively by the RNA Society for the first 12 months after the full-issue publication date (see http://rnajournal.cshlp.org/site/misc/terms.xhtml). After 12 months, it is available under a Creative Commons License (Attribution-NonCommercial 4.0 International), as described at http://creativecommons.org/licenses/by-nc/4.0/.

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FIGURE 2. — Full-length SF3A1-UBL domain is required for RNA binding. (A) Alignment of the SF3A1 UBL domain (aa 703–793) to ubiquitin. Graphical representations of secondary structure in relation to primary structure were created based on PDB entries for ubiquitin (PDB ID: 1UBQ) and SF3A1-UBL (PDB ID: 1ZKH). In the alignment, a dot indicates the presence of an identical residue to the reference sequence used in the alignment (ubiquitin), and a tilde indicates a gap. (B) Schematic of all amino- and carboxy-terminal deletion constructs made to study the SF3A1-UBL domain. (C) Western blot of 6×His-tagged proteins in CFE reactions containing the EV control, or expression vectors for FL (UBL_703–793), amino-terminal truncated (UBL_714–793), and carboxy-terminal truncated (UBL_703–790 and UBL_703–786) SF3A1-UBL. The expressed UBL domain proteins were detected with anti-6×His primary antibody. (D) UV-crosslinking of ³²P-U1-SL4 in CFE reactions expressing EV or UBL proteins. Arrows indicate specific crosslinked products, and asterisks indicate nonspecific products that were also observed in reactions containing the EV control.