Fig. 5: Evolutionary constraints on SLY1 reversion routes.
From: Evolution of a plant growth-regulatory protein interaction specificity
a, Alignment of amino acid sequences of SLY1 orthologues. Black highlights identity, grey highlights residue similarity, red arrows indicate sites in AtSLY1 targeted by selected amino acid substitutions (and equivalent sites in MpSLY1, SmSLY1 and OsGID2). For more comprehensive alignment, see Extended Data Fig. 7. b, Interactions between MpSLY1 or OsGID2 (or indicated variants) and MpDELLA or OsSLR1. c, Mean (±s.d.) strengths of yeast 2-hybrid interactions between AtSLY1 (or indicated variants) or MpSLY1 (or indicated variants) and GAI. Red dots indicate individual values (n = 3), different letters (a–d) indicate significant differences (one-way ANOVA with Tukey’s test). d, Mean (±s.d.) strengths of yeast 2-hybrid interactions between AtSLY1 (or indicated variants) and GAI. Red dots indicate individual values (n = 3), different letters (a–g) indicate significant differences (one-way ANOVA with Tukey’s test). e, Comparison of sites where amino acid substitutions confer increased affinity for the DELLA A form. Data from EP-PCR/yeast 2-hybrid screens. For SmSLY1, mutants were initially selected for increased affinity for GAI, then validated with SmDELLA1. For AtSLY1, substitutions were as previously described (Fig. 2a). For OsGID2, mutants were initially selected for increased affinity for GAI, then validated with the rice DELLA SLR1. Targeted amino acids occupying equivalent positions are indicated with the same colour. Specifically, SmSLY1 E91 is equivalent to OsGID2 L136 (orange); SmSLY1 L102 is equivalent to OsGID2 V147 (blue); SmSLY1 L139 is equivalent to AtSLY1 E138 (purple); SmSLY1 S129 is equivalent to AtSLY1 T128 and OsGID2 S180 (red). All other targeted positions (black) are unique to SmSLY1 or AtSLY1. f, Interaction of indicated OsGID2 and Osgid2 variants with SLR1.
