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. 2018 Sep;24(9):1214–1228. doi: 10.1261/rna.066373.118

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© 2018 Black 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. — The N terminus of Utp14 interacts with an extraribosomal Rps7-Rps22 heterodimer. (A) Truncations from either termini of Utp14 are unable to support growth as shown by 10-fold serial dilutions of GAL1::3xHA-UTP14 cells (AJY3243) harboring vectors encoding UTP14-TAP (pAJ4176), utp14-ΔC-TAP (pAJ4177), utp14-ΔN-TAP (pAJ4178), or empty vector (pRS415) spotted on media lacking leucine containing either galactose or glucose. (B) Western blot analysis of sucrose gradient fractions showing that the different Utp14 truncation mutants sediment into the gradients. (C) Coomassie-stained gel of proteins that copurified with full-length or truncated Utp14 proteins. Pellet and supernatant fractions were separated by overlaying eluate onto sucrose cushions followed by ultracentrifugation. The arrow heads in lane 6 indicate Utp14-ΔC (∼150 kDa), Rps7 (∼20 kDa), and Rps22 (∼10 kDa). (D) Yeast two-hybrid interaction data for Rps7 and Rps22. (E) Yeast two-hybrid interaction between Utp14 and Rps7 are shown. A cartoon of the Utp14 constructs is shown to the right. (Abbreviations as in legend of Figure 2.)