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. 2018 Aug;24(8):1028–1040. doi: 10.1261/rna.065664.118

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© 2018 Carrocci 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 4. — Humanized Hsh155 functions in yeast and alters nonconsensus BS usage. (A) In vitro splicing assay with WT Hsh155 in the presence or absence of PB (20 µM). PB does not inhibit yeast splicing at this concentration. (B) Schematic illustration of the humanized Hsh155 chimeric proteins. (C,D) Growth of haploid yeast containing the indicated humanized Hsh155 proteins on nonselective (−Trp) and selective (−Trp +FOA) media at 30°C. (E) Heatmap summarizing ACT1-CUP1 reporter data for the indicated humanized Hsh155 proteins and MDS mutants with several BS reporters. Plotted data represent the log₂ transform of the ratio of the maximum [Cu²⁺] at which growth was observed for the indicated mutant to the maximum [Cu²⁺] at which growth was observed for WT Hsh155. Purple indicates decreased growth relative to WT, and orange indicates improved growth. (F) Heatmap of ACT1-CUP1 reporter data as in panel E but including the Hs5-16 humanized protein with and without the R1074H splicing inhibitor resistance mutation (note that numbering refers to human SF3b1).